Fusion genes associated with acute megakaryoblastoc leukemias

ABSTRACT

The invention relates to human nucleotide sequences which occur as a result of the t(1;22)(p13;q13) chromosomal translocation event which is known to occur almost invariable in young children with acute megakaryoblastic leukemia. The translocation results in the formation of fusion genes which encode fusion proteins. The invention provides the nucleotide sequences of transcripts of the fusion genes and the amino acid sequences of the fusion proteins encoded thereby. Also provided are methods for detecting the t(1, 22) translocation, for identifying agents capable of binding to the fusion protein and for identifying agents useful for treating patients with acute megakaryoblastic leukemia.

FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0001] This invention was made in part with U.S. Government supportunder Cancer Center Support (CORE) grants CA-21765 and CA-87064 from theNational Cancer Institute. The Government may have certain rights inthis invention.

FIELD OF THE INVENTION

[0002] The present invention is directed to the field of the moleculargenetics of cancer. Specifically, the present invention relates to humanacute megakaryoblastic leukemias in which a translocation betweenchromosomes 1 and 22 (referred to in the art as “t(1 ;22)”) hasoccurred. On a molecular level, the DNA rearrangement in t(1 ;22)results in two reciprocal fusion genes that each comprise segments ofthe RBM15 and MKL1 genes.

BACKGROUND OF THE INVENTION

[0003] Chromosomal abnormalities are frequently associated withmalignant diseases. In a number of instances, specific chromosomaltranslocations have been characterized, which generate fusion genesencoding proteins with oncogenic properties (Sawyers et al., Cell64:337-350 (1991)).

[0004] Recently, the cloning of chromosomal translocations has led toidentification of pathogenically relevant oncogenic fusion transcriptsand proteins in specific subsets of acute nonlymphocytic leukemia(ANLL), such as promyelocytic leukemia-retinoic acid receptor alphafusion gene (PML-RARα) in acute promyelocytic leukemia (FAB-M3 subtype),the acute myeloid leukemia 1-eight twenty one fusion gene (AML1-ETO) inANLL with maturation (FAB-M2), and various mixed lineage leukemia (MLL)fusions in acute myelomonocytic and monocytic leukemias (FAB-M4 and -M5)(Melnick, A. & Licht, J. D., Blood 93, 3167-3215 (1999); Downing, J. R.,Br. J Haematol. 106, 296-308 (1999); Rowley, J. D., Semin. Hematol. 36,59-72 (1999); Look, A. T., Science 278, 1059-1064 (1997); Faretta, M.,Di Croce, L. & Pelicci, P. G., Sem. Hematol. 38, 42-53 (2001)). Despitethese significant advances, little is known about the genetic mechanismsunderlying acute leukemias of the megakaryoblastic (platelet precursor)lineage (AMKL, FAB-M7) (Cripe, L. D, infra). Almost invariably, AMKL innon-Down syndrome infants and young children harbor the t(1;22)(p13;q13)translocation, in most cases as the sole cytogenetic abnormality(Carroll, A. et al; Lion, T. et al., and Bernstein, J. et al., infra).Phenotypically, AMKL presents de novo (i.e., without a so-calledpreleukemic stage), with a large leukemia cell mass, and frequentfibrosis of bone marrow and other organs. Progression is usually rapiddespite therapy, with a median overall patient survival of only eightmonths. Thus, compositions and methods for the early and accuratediagnosis and treatment of leukemia, particularly AMKL, are needed.

SUMMARY OF THE INVENTION

[0005] Compositions and methods associated with the diagnosis andtreatment of leukemia are provided. The invention discloses theidentification, cloning and sequencing of human nucleotide sequencescorresponding to the t(1;22)(p13;q13) chromosomal translocation eventwhich occurs in individuals with acute megakaryoblastic leukemia (AMKL).The rearrangement recombines sequences from the RNA-bindingmotifprotein-15 gene (RBM15) on chromosome 1p13 with those from theMegakaryoblastic Leukemia-1 gene (MKL1) on chromosome 22q13. As a resultof the t(1;22)(p13;q13) rearrangement, two fusion genes, one on each ofthe two derivative chromosomes (der(1) and der(22)), are produced. Thefirst fusion gene, designated RBM15-MKL1, resides on der(22) andcomprises a 5′ portion of the RBM15 and a 3′ segment of the MKL1. Thesecond fusion gene, designated as MKL1-RBM15, resides on der(1) andcomprises a 5′ portion of the MKL1 and a 3′ segment of the RBM15. Bothfusion genes are transcribed. A single transcript is expressed fromRBM15-MKL1. Two transcripts are found to be expressed from MKL1-RBM15.Isolated nucleotide molecules comprising the nucleotide sequences of theRBM15-MKL1 gene transcript and the two MKL1-RBM15 gene transcripts,MKL1-RBM15s and MKL1-RBM15_(S+AE) are provided. Additionally providedare the amino acid sequences of the fusion proteins encoded by theRBM15-MKL1 transcript and the two MKL1-RBM15 transcripts.

[0006] Utilizing the sequences of the present invention, the presentinvention provides methods of identifying the presence of nucleic acidscontaining the RBM15-MKL1 fusion gene and/or MKL1-RBM15 fusion gene andthe transcripts of these fusion genes in a sample. Such methods can beused in diagnosis and treatment, for example, to determine if particularcells or tissues express RBM15-MKL1 or MKL1-RBM15 coding sequences, ordiagnostic assays to determine if a mammal has leukemia or a geneticpredisposition to (i.e., is at an increased risk of developing)leukemia.

[0007] The RBM15-MKL1 and MKL1-RBM15 fusion proteins and polypeptidesequences of the invention, whether produced by host/vector systems orotherwise, can be used to produce antibodies which specificallyrecognize (i.e., bind) the RBM15-MKL1 and MKL1-RBM15 fusion proteins,respectively. The invention provides methods of identifying the presenceof nucleic acids encoding the RBM15-MKL1 fusion protein and/orMKL1-RBM15 fusion protein in a sample involving the use of suchantibodies. Such methods find use in diagnosis and treatment of AMKL,for example, to determine if particular cells or tissues express theRBM15-MKL1 fusion protein and/or the MKL1-RBM15 fusion proteins and toinhibit the activity of these fusion proteins.

[0008] The present invention also provides for transgenic cells andanimals, preferably mice, which (a) contain and express an RBM15-MKL1fusion gene derived from an exogenous source and subsequently introducedinto the genome of the cell or animal, (b) contain and express a geneencoding an RBM15-MKL1 fusion protein derived from an exogenous sourceand subsequently introduced into the genome of the cell or animal, (c)contain and express an MKL1-RBM15 fusion gene derived from an exogenoussource and subsequently introduced into the genome of the cell or animal(d) contain and express a gene encoding an MKL1-RBM15 fusion proteinderived from an exogenous source and subsequently introduced into thegenome of the cell or animal, (e) contain and express both a geneencoding an MKL1-RBM15 fusion protein and a gene encoding an RBM15-MKL1fusion protein, with both genes derived from an exogenous source andsubsequently introduced into the genome of the animal, (f) knock-out theexpression of the RBM15 and/or MKL1 genes, or (g) knock-out theexpression of the RBM15-MKL1 and/or MKL1-RBM15 fusion proteins in anAMKL cell line. Methods of utilizing such cells and mice to identify andtest carcinogenic or therapeutic compositions are also described herein.

[0009] The nucleotide sequences of the RBM15-MKL1 and MKL1-RBM15 genesand the coding sequences of the RBM15-MKL1 and the two MKL1-RBM15 fusionproteins of the invention can also be utilized to design and prepareagents which specifically inhibit the expression of the RBM15-MKL1 orMKL1-RBM15 genes in cells for therapeutic and other purposes. TheRBM15-MKL1 and MKL1-RBM15 nucleotide sequences of the invention can befurther utilized in methods of producing the RBM15-MKL1 and MKL1-RBM15fusion proteins, respectively.

[0010] The present invention further provides methods for isolating andidentifying the natural ligand(s) and gene targets bound by theRBM15-MKL1 and MKL1-RBM15 fusion proteins, and for identifyingderivatives of the ligand(s) or synthetic compounds that act to inhibitthe action of the RBM15-MKL1 and MKL1-RBM15 fusion proteins.

[0011] Additionally provided are compartmentalized kits to receive inclose confinement one or more containers containing the reagents used inone or more of the above described detection methods.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012]FIG. 1 is a schematic representation of the normal RBM15 and MKL1proteins, and the fusion proteins formed by t(1;22). The locations ofthe fusion junctions in the proteins are indicated by vertical arrows.The 931-aa MKL1 protein (predicted mass, 98.9 kDa; SEQ ID. NO: 14)contains a bipartite nuclear localization signal (N, residues 14-31 ofSEQ ID NO: 14), a single SAP DNA-binding motif (residues 347-381 of SEQID NO: 14), a coiled-coil region (CC, residues 521-563 of SEQ ID NO:14), and a long C-terminal proline-rich segment (P, residues 564-811 ofSEQ ID NO: 14). The three isofomis of RBM15 (RBM15_(S), RBM15_(S+AE) andRBM15_(L)) share an identical 2863-bp 5′ coding sequence (nucleotides 84to 2946 of SEQ ID Nos. 7, 9, and 11) and differ only in their extremeC-termini, distal to the location of the t(1;22) fusion junction, due tosplicing of alternative exons. Specifically, RBM15_(L), the longestRBM15 transcript (approx. 9 kb), contains a unique 416-bp 3′ exon(nucleotides 2947 to 3362 of SEQ ID NO: 11) and encodes a predicted957-aa protein (mass, 104.6 kDa; SEQ ID NO: 12), whereas the shortesttranscript, RBM15_(S) (approx. 4 kb), possesses a divergent 3′ sequence(nucleotides 2947to 3312 of SEQ ID NO: 7) and encodes a 969-aa protein(mass, 106.2 kDa; SEQ ID NO: 8). Detailed analysis of the approximately4-kb RBM15 transcript revealed a variant, RBM15_(S+AE) (short transcriptplus alternative exon), which contains an additional 111-bp exon(nucleotides 2947 to 3057of SEQ ID NO: 9) interposed between the2,863-bp common sequence and the 366-bp sequence in RBM₁₅ _(S) andencodes a polypeptide of 977 aa (mass, 107.1 kDa; SEQ ID NO: 10). AllRBM15 isoforms contain three RNA recognition motifs (RRM; residues170-252, 374-451 and 455-529 of SEQ ID Nos. 8, 10, and 12), a bipartitenuclear localization signal (N, residues 716-733 of SEQ ID Nos. 8, 10,and 12), and a SPOC domain (S, residues 714-954 of SEQ ID Nos. 8, 10,and 12). Several regions in RBM15 of potential functional importance arecharacterized by a high content of specific aa, including aglycine/serine-rich segment (residues 60-166 of SEQ ID Nos. 8, 10, and12), a small proline-rich (LPPPPPPPLP) motif (residues 315-324 of SEQ IDNos. 8, 10, and 12), an arginine-rich portion (residues 616-732 of SEQID Nos. 8, 10, and 12), and a short C-terminal serine-rich segment(GSSDSRSSSSSAASD) at amino acids 865-879 of SEQ ID Nos. 8, 10, and 12.The 1883-aa RBM15-MKL1 chimeric protein (mass, 203.1 kDa; SEQ ID NO: 2)is comprised of the common N-terminal portion of RBM15 (residues 1 to954 of SEQ ID Nos. 8, 10, and 12) and all but the first two residues ofMKL1, thus containing all identified motifs of each normal protein. Bycontrast, the reciprocal MKL1-RBM15 fusions contain only the first twoaa of MKL1 fused to the short C-terminal sequences of either RBM15_(S)or RBM15_(S+AE), and are predicted to encode peptides of only 17 (SEQ IDNO: 4) and 25 (SEQ ID NO: 6) amino acids, respectively. The portions ofthe schematic illustrating the alternative C-termini of RBM15 and thepredicted MKL1-RBM15 fusion peptides are enlarged for clarity, and arethus not to scale.

DETAILED DESCRIPTION OF THE INVENTION

[0013] Compositions and methods for the identification, diagnosis, andtreatment of leukemia or a genetic predisposition to leukemia areprovided. The present invention is based on the discovery of tworeciprocal fusion genes, RBM15-MKL1 and MKL1-RBM15, that result from thet(1;22)(p13;q13) chromosomal translocation event associated with acutemegakaryoblastic leukemia (AMKL). In particular, the invention providesthe novel nucleotide sequences of a transcript of the RBM15-MKL1 fusiongene (SEQ ID NO: 1) and two transcripts, MKL1-RBM15_(S) (SEQ ID NO: 3)and MKL1-RBM15_(S+AE) (SEQ ID NO: 5), of the MKL1-RBM15 fusion gene.Additionally provided are the amino acid sequences (SEQ ID NOS: 2, 4,and 6, respectively) of the fusion proteins encoded by such nucleotidesequences. Such nucleotide sequences and amino acid sequences find use,for example, in methods for detecting the t(1;22)(p13;q13) chromosomaltranslocation event associated with AMKL, methods for identifying agentsthat bind to the fusion proteins and methods for identifying agentsuseful for treating AMKL.

[0014] In addition to the RBM15-MKL1 and MKL1-RBM15 nucleotide and aminoacid sequences, the invention provides isolated nucleotide moleculescomprising nucleotide sequences of three transcripts of the RBM15 gene,RBM15_(S) (SEQ ID NO: 7), RBM15_(S+AE) (SEQ ID NO: 9), and RBM15_(L)(SEQ ID NO: 11) and the nucleotide sequence of MKL1 (SEQ ID NO: 13).Further provided are isolated proteins comprising the amino acidssequences of the proteins encoded thereby, RBM15_(S) (SEQ ID NO: 8),RBM15_(s+AE) (SEQ ID NO: 10), and RBM15_(L) (SEQ ID NO: 12) and MKL1(SEQ ID NO: 14) respectively. Such nucleotide and amino acid sequencesfind use, for example, in methods for detecting the t(1;22)(p13;q13)chromosomal translocation event associated with AMKL and methods foridentifying agents useful for treating AMKL.

[0015] The RBM15 and MKL1 proteins, as well as the noted fusion proteinsderived from RBM15 and MKL1, are contemplated to act as transcriptionfactors which bind to corresponding DNA regulatory sequences. Thus theseproteins may be used to regulate the expression of genes that includethe regulatory DNA sequences that these proteins recognize. Methods foridentifying such regulatory DNA sequences based on their ability to bindRBM15, MKL1, RBM15-MKL1 and MKL1-RBM15 are also included in the presentinvention.

[0016] The nucleotide and amino acid sequences of the invention are setforth in the sequence listing. Below is a brief description of thesequences in the sequences listing.

[0017] SEQ ID NO: 1 is the nucleotide sequence RBM15-MKL1 cDNA. The openreading frame is from nucleotide 84 through nucleotide 5732.

[0018] SEQ ID NO: 2 is the amino acid sequence of the RBM15-MKL1 fusionprotein.

[0019] SEQ ID NO: 3 is the nucleotide sequence MKL1-RBM15_(S) cDNA. Theopen reading frame is from nucleotide 551 through nucleotide 601.

[0020] SEQ ID NO: 4 is the amino acid sequence of the MKL1 -RBM15_(S)fusion protein.

[0021] SEQ ID NO: 5 is the nucleotide sequence MKL1-RBM15_(S+AE) cDNA.The open reading frame is from nucleotide 551 through nucleotide 625.

[0022] SEQ ID NO: 6 is the amino acid sequence of the MKL1-RBM15_(S+AE)fusion protein.

[0023] SEQ ID NO: 7 is the nucleotide sequence RBM15_(S) cDNA. The openreading frame is from nucleotide 84 through nucleotide 2990.

[0024] SEQ ID NO: 8 is the amino acid sequence of the RBM15_(S) protein.

[0025] SEQ ID NO: 9 is the nucleotide sequence RBM15_(S+AE) cDNA. Theopen reading frame is from nucleotide 84 through nucleotide 3014.

[0026] SEQ ID NO: 10 is the amino acid sequence of the RBM15_(S+AE)protein.

[0027] SEQ ID NO: 11 is the nucleotide sequence RBM15_(L) cDNA. The openreading frame is from nucleotide 84 through nucleotide 2954.

[0028] SEQ ID NO: 12 is the amino acid sequence of the RBM15_(L)protein.

[0029] SEQ ID NO: 13 is the nucleotide sequence MKL1 cDNA. The openreading frame is from nucleotide 551 through nucleotide 3346.

[0030] SEQ ID NO: 14 is the amino acid sequence of the MKL1 protein.

[0031] SEQ ID NO: 15 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1-294R.

[0032] SEQ ID NO: 16 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1-73R.

[0033] SEQ ID NO: 17 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1 -59R.

[0034] SEQ ID NO: 18 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15(S)-2746F.

[0035] SEQ ID NO: 19 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1-204R.

[0036] SEQ ID NO: 20 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1-F.

[0037] SEQ ID NO: 21 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15(S)-2930R.

[0038] SEQ ID NO: 22 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15(L)-1636R.

[0039] SEQ ID NO: 23 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-1118F.

[0040] SEQ ID NO: 24 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-1551R.

[0041] SEQ ID NO: 25 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-2831F.

[0042] SEQ ID NO: 26 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-3149R.

[0043] SEQ ID NO: 27 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-1616F.

[0044] SEQ ID NO: 28 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated RBM15-2004R.

[0045] SEQ ID NO: 29 is the nucleotide sequence of the syntheticoligonucleotide primer herein designated MKL1-155F.

[0046] Based on these observations, one embodiment of the presentinvention provides a first isolated nucleotide molecule comprising thecoding sequence (SEQ ID NO: 1) of the RBM15-MKL1 fusion protein, whichencodes the RBM15-MKL1 fusion protein (SEQ ID NO: 2), a second isolatednucleotide molecule, comprising the coding sequence (SEQ ID NO: 3) ofthe MKL1-RBM15_(S) fusion protein, which encodes the MKL1-RBM15_(S)fusion protein (SEQ ID NO: 4), a third isolated nucleotide moleculecomprising the coding sequence (SEQ ID NO: 5) of the MKL1-RBM15_(S+AE)fusion protein which encodes the MKL1-RBM15_(S+AE) fusion protein (SEQID NO: 6), a fourth isolated nucleotide molecule, comprising the codingsequence (SEQ ID NO: 7) of the RBM15_(S) protein, which encodes theRBM15_(S) protein (SEQ ID NO: 8), a fifth isolated nucleotide moleculecomprising the coding sequence (SEQ ID NO: 9) of the RBM15_(S+AE)protein which encodes the RBM15_(S+AE) protein (SEQ ID NO: 10), a sixthisolated nucleotide molecule, comprising the coding sequence (SEQ ID NO:11) of the RBM15_(L) protein, which encodes the RBM15L protein (SEQ IDNO: 12), and a seventh isolated nucleotide molecule comprising thecoding sequence (SEQ ID NO: 13) of the MKL1 protein, which encodes theMKL1 protein (SEQ ID NO: 14).

[0047] It is recognized that nucleotide molecules and proteins of theinvention will have a nucleotide or an amino acid sequence sufficientlyidentical to a nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, or13 or to an amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, or 14.The term “sufficiently identical” is used herein to refer to a firstamino acid or nucleotide sequence that contains a sufficient or minimumnumber of identical or equivalent (e.g., with a similar side chain)amino acid residues or nucleotides to a second amino acid or nucleotidesequence such that the first and second amino acid or nucleotidesequences have a common structural domain and/or common functionalactivity. For example, amino acid or nucleotide sequences that contain acommon structural domain having at least about 45%, 55%, or 65%identity, preferably 75% identity, more preferably 85%, 95%, or 98%identity are defined herein as sufficiently identical.

[0048] To determine the percent identity of two amino acid sequences orof two nucleic acids, the sequences are aligned for optimal comparisonpurposes. The percent identity between the two sequences is a functionof the number of identical positions shared by the sequences (i.e.,percent identity=number of identical positions/total number of positions(e.g., overlapping positions)×100). In one embodiment, the two sequencesare the same length. The percent identity between two sequences can bedetermined using techniques similar to those described below, with orwithout allowing gaps. In calculating percent identity, typically exactmatches are counted.

[0049] The determination of percent identity between two sequences canbe accomplished using a mathematical algorithm. A preferred, nonlimitingexample of a mathematical algorithm utilized for the comparison of twosequences is the algorithm of Karlin and Altschul (1990) Proc. Natl.Acad Sci. USA 87:2264, modified as in Karlin and Altschul (1993) Proc.Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm is incorporatedinto the NBLAST and XBLAST programs of Altschul et al. (1990) J. Mol.Biol. 215:403. BLAST nucleotide searches can be performed with theNBLAST program, score=100, wordlength=12, to obtain nucleotide sequenceshomologous to the nucleic acid molecules of the invention. BLAST proteinsearches can be performed with the XBLAST program, score=50,wordlength=3, to obtain amino acid sequences homologous to proteinmolecules of the invention. To obtain gapped alignments for comparisonpurposes, Gapped BLAST can be utilized as described in Altschul el al.(1997) Nucleic Acids Res. 25:3389. Alternatively, PSI-Blast can be usedto perform an iterated search that detects distant relationships betweenmolecules. See Altschul et al. (1997) supra. When utilizing BLAST,Gapped BLAST, and PSI-Blast programs, the default parameters of therespective programs (e.g., XBLAST and NBLAST) can be used. Seehttp://www.ncbi.nlm.nih.gov. Another preferred, non-limiting example ofa mathematical algorithm utilized for the comparison of sequences is thealgorithm of Myers and Miller (1988) CABIOS 4:11-17. Such an algorithmis incorporated into the ALIGN program (version 2.0), which is part ofthe GCG sequence alignment software package. When utilizing the ALIGNprogram for comparing amino acid sequences, a PAM120 weight residuetable, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0050] The invention encompasses RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1 nucleicacid molecules and fragments thereof. Nucleic acid molecules that arefragments of these nucleotide sequences are also encompassed by thepresent invention. By “fragment” is intended a portion of the nucleotidesequence encoding an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L) or MKL1 protein. A fragment of anRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L) or MKL1 nucleotide sequence may encode a biologically activeportion of an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L) or MKL1 protein, or it may be a fragment thatcan be used as a hybridization probe or PCR primer using methodsdisclosed below. A biologically active portion of an RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L) orMKL1 protein can be prepared by isolating a portion of one of theRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), and MKL1 nucleotide sequences of the invention, expressingthe encoded portion of the RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L) or MKL1 protein(e.g., by recombinant expression in vitro), and assessing the activityof the encoded portion of the RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1 protein.Nucleic acid molecules that are fragments of an RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L),and MKL1 nucleotide sequence comprise at least about 15, 20, 50, 75,100, 200, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850,900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1500,1600, 1700, 1800, 1900, 2000, 2500, 3000, 3500, 4500, 5000, 5500, 6000,or 6500 nucleotides, or up to the number of nucleotides present in afull-length nucleotide sequence disclosed herein (for example, 6836,923, 1034, 3312, 3423, 3383, and 4447 nucleotides for SEQ ID NOS: 1, 3,5, 7, 9, 11, and 13, respectively) depending upon the intended use.

[0051] It is understood that isolated fragments include any contiguoussequence not disclosed prior to the invention as well as sequences thatare substantially the same and which are not disclosed. Accordingly, ifan isolated fragment is disclosed prior to the present invention, thatfragment is not intended to be encompassed by the invention. When asequence is not disclosed prior to the present invention, an isolatednucleic acid fragment is at least about 12, 15, 20, 25, or 30 contiguousnucleotides. Other regions of the nucleotide sequence may comprisefragments of various sizes, depending upon potential homology withpreviously disclosed sequences.

[0052] A fragment of an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L) or MKL1nucleotide sequence thatencodes a biologically active portion of an RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 proteinof the invention will encode at least about 15, 25, 30, 50, 75, 100,125, 150, 175, 200, 250, or 300 contiguous amino acids, or up to thetotal number of amino acids present in a full-length RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L),or MKL1protein of the invention (for example, 1883, 17, 25, 969, 977,957, and 931 amino acids for SEQ ID NOS: 2, 4, 6, 8, 10, 12, and 14,respectively). Fragments of an RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L) and MKL1nucleotidesequence that are useful as hybridization probes for PCR primersgenerally need not encode a biologically active portion of anRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), and MKL1 protein, respectively.

[0053] Nucleic acid molecules that are variants of the nucleotidesequences disclosed herein are also encompassed by the presentinvention. “Variants” of the RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1nucleotide sequences include those sequences that encode the RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L),and MKL1 proteins, respectively, disclosed herein but that differconservatively because of the degeneracy of the genetic code. Thesenaturally occurring allelic variants can be identified with the use ofwell-known molecular biology techniques, such as polymerase chainreaction (PCR) and hybridization techniques as outlined below. Variantnucleotide sequences also include synthetically derived nucleotidesequences that have been generated, for example, by using site-directedmutagenesis but which still encode the RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 proteindisclosed in the present invention as discussed below. Generally,nucleotide sequence variants of the invention will have at least about45%, 55%, 65%, 75%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97, 98%, or99% identity to a particular nucleotide sequence disclosed herein. Avariant RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L) or MKL1 nucleotide sequence will encode avariant RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L), or MKL1 protein, respectively, that has anamino acid sequence having at least about 45%, 55%, 65%, 75%, 85%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97, 98%, or 99% identity to the amino acidsequence of an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L), or MKL1 protein disclosed herein.

[0054] In addition to the RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L) and MKL1 nucleotide sequences shownin SEQ ID NOS: 1, 3, 5, 7, 9, 11, and 13 , it will be appreciated bythose skilled in the art that DNA sequence polymorphisms that lead tochanges in the amino acid sequences of RBM5-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 proteinsmay exist within a population (e.g., the human population). Such geneticpolymorphism in an RBM15-MKL1, MKL1-RBM15, RBM15, and MKL1 gene mayexist among individuals within a population due to natural allelicvariation. An allele is one of a group of genes that occur alternativelyat a given genetic locus. As used herein, the terms “gene” and“recombinant gene” refer to nucleic acid molecules comprising an openreading frame encoding an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 protein, preferably amammalian RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM¹⁵ _(S+AE), RBM15_(L), or MKL1 protein. As used herein, the phrase“allelic variant” refers to a nucleotide sequence that occurs at anRBM15-MKL1, MKL1-RBM15, RBM15, and MKL1 locus or to a polypeptideencoded by the nucleotide sequence. Such natural allelic variations cantypically result in 1-5% variance in the nucleotide sequence of theRBM15-MKL1, MKL1-RBM15, RBM15, and MKL1 gene. Any and all suchnucleotide variations and resulting amino acid polymorphisms orvariations in an RBM15-MKL1, MKL1-RBM15, RBM15, and MKL1 amino acidsequence that are the result of natural allelic variation and that donot alter the functional activity of RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1 proteinsare intended to be within the scope of the invention.

[0055] Moreover, nucleic acid molecules encoding RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L),and MKL1 proteins from other species (RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1homologues), which have a nucleotide sequence differing from that of theRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), and MKL1 sequences disclosed herein, are intended to bewithin the scope of the invention. For example, nucleic acid moleculescorresponding to natural allelic variants and homologues of the humanRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L) and MKL1 cDNAs of the invention can be isolated based on theiridentity to the human RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1 nucleic acids disclosedherein using the human cDNA, or a portion thereof, as a hybridizationprobe according to standard hybridization techniques under stringenthybridization conditions as disclosed herein.

[0056] In addition to naturally occurring allelic variants of theRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), and MKL1 sequences that may exist in the population, theskilled artisan will further appreciate that changes can be introducedby mutation into the nucleotide sequences of the invention therebyleading to changes in the amino acid sequence of the encoded RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L),and MKL1 proteins, respectively, without altering the biologicalactivity of the RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1 proteins. Thus, an isolatednucleic acid molecule encoding an RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 proteinhaving a sequence that differs from that of SEQ ID NOS: 2, 4, 6, 8, 10,12, or 14, respectively, can be created by introducing one or morenucleotide substitutions, additions, or deletions into the correspondingnucleotide sequence disclosed herein, such that one or more amino acidsubstitutions, additions or deletions are introduced into the encodedprotein. Mutations can be introduced by standard techniques, such assite-directed mutagenesis and PCR-mediated mutagenesis. Such variantnucleotide sequences are also encompassed by the present invention.

[0057] For example, preferably, conservative amino acid substitutionsmay be made at one or more predicted, preferably nonessential amino acidresidues. A “nonessential” amino acid residue is a residue that can bealtered from the wild-type sequence of an RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 protein(e.g., the sequence of SEQ ID NOS: 2, 4, 6, 8, 10, 12, or 14,respectively) without altering the biological activity, whereas an“essential” amino acid residue is required for biological activity. A“conservative amino acid substitution” is one in which the amino acidresidue is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art. These families include amino acids with basicside chains (e.g., lysine, arginine, histidine), acidic side chains(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine),nonpolar side chains (e.g., alanine, valine, leucine, isoleucine,proline, phenylalanine, methionine, tryptophan), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine). Suchsubstitutions would not be made for conserved amino acid residues, orfor amino acid residues residing within a conserved motif.

[0058] Alternatively, variant RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1nucleotide sequences can be made by introducing mutations randomly alongall or part of an RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L) or MKL1 coding sequence, such as bysaturation mutagenesis, and the resultant mutants can be screened forRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), or MKL1 biological activity to identify mutants that retainactivity. Following mutagenesis, the encoded protein can be expressedrecombinantly, and the activity of the protein can be determined usingstandard assay techniques.

[0059] Thus, the nucleotide sequences of the invention include thesequences disclosed herein as well as fragments and variants thereof.The RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L), and MKL1 nucleotide sequences of the invention,and fragments and variants thereof, can be used as probes and/or primersto identify and/or clone RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15, and MKL1 homologues in other cell types,e.g., from other tissues, as well as homologues from other mammals. Suchprobes can be used to detect transcripts or genomic sequences encodingthe same or identical proteins.

[0060] In this manner, methods such as PCR, hybridization, and the likecan be used to identify such sequences having substantial identity tothe sequences of the invention. See, for example, Sambrook et al. (1989)Molecular Cloning: Laboratory Manual (2d ed., Cold Spring HarborLaboratory Press, Plainview, N.Y.) and Innis, et al. (1990) PCRProtocols: A Guide to Methods and Applications (Academic Press, NY).RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), and MKL1 nucleotide sequences isolated based on theirsequence identity to the RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L) and MKL1 nucleotide sequences setforth herein or to fragments and variants thereof are encompassed by thepresent invention.

[0061] In a hybridization method, all or part of a known RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L) orMKL1nucleotide sequence can be used to screen cDNA or genomic libraries.Methods for construction of such cDNA and genomic libraries aregenerally known in the art and are disclosed in Sambrook et al. (1989)Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring HarborLaboratory Press, Plainview, N.Y.). The so-called hybridization probesmay be genomic DNA fragments, cDNA fragments, RNA fragments, or otheroligonucleotides, and may be labeled with a detectable group such as³²P, or any other detectable marker, such as other radioisotopes, afluorescent compound, an enzyme, or an enzyme co-factor. Probes forhybridization can be made by labeling synthetic oligonucleotides basedon the known RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L), and MKL1 nucleotide sequence disclosed herein.Degenerate primers designed on the basis of conserved nucleotides oramino acid residues in a known RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM₁₅ _(S), RBM15_(S+AE), RBM15_(L) and MKL1nucleotide sequence or encoded amino acid sequence can additionally beused. The probe typically comprises a region of nucleotide sequence thathybridizes under stringent conditions to at least about 12, preferablyabout 25, more preferably about 50, 75, 100, 125, 150, 175, 200, 250,300, 350, or 400 consecutive nucleotides of an RBM15-MKL1,MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15, RBM15_(S+AE), RBM15 and MKL1nucleotide sequence of the invention or a fragment or variant thereof.Preparation of probes for hybridization is generally known in the artand is disclosed in Sambrook et al. (1989) Molecular Cloning: ALaboratory Manual (2d ed., Cold Spring Harbor Laboratory Press,Plainview, N.Y.), herein incorporated by reference.

[0062] As used herein, the term “hybridizes under stringent conditions”is intended to describe conditions for hybridization and washing underwhich nucleotide sequences having at least about 60%, 65%, 70%,preferably 75% identity to each other typically remain hybridized toeach other. Such stringent conditions are known to those skilled in theart and can be found in Current Protocols in Molecular Biology (JohnWiley & Sons, New York (1989)), 6.3.1-6.3.6. A preferred, non-limitingexample of stringent hybridization conditions is hybridization in 6×sodium chloride/sodium citrate (SSC) at about 45° C., followed by one ormore washes in 0.2×SSC, 0.1% SDS at 50-65° C. In another preferredembodiment, stringent conditions comprise hybridization in 6×SSC at 42°C., followed by washing with 1×SSC at 55° C. Preferably, an isolatednucleic acid molecule that hybridizes under stringent conditions to anRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM51,S or MKL1 nucleotide sequence of the invention corresponds to anaturally occurring nucleic acid molecule. As used herein, a “naturallyoccurring” nucleic acid molecule refers to an RNA or DNA molecule havinga nucleotide sequence that occurs in nature (e.g., encodes a naturalprotein).

[0063] Thus, in addition to the RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(S), and MKL1nucleotide sequences disclosed herein and fragments and variantsthereof, the isolated nucleic acid molecules of the invention alsoencompass homologous DNA sequences identified and isolated from othercells and/or organisms by hybridization with entire or partial sequencesobtained from the RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE),RBM15_(S), RBM15_(S+AE), RBM15_(L), and MKL1. By inserting any of thenucleotide sequences of the present invention into an appropriatevector, one skilled in the art can readily produce large quantities ofthe specific sequence. Alternatively, the RBM15-MKL1, MKL1-RBM15_(S) andMKL1-RBM15_(S+AE), nucleotide sequences of the invention can be furtherutilized in methods of producing the RBM15-MKL1, MKL1-RBM15_(S) andMKL1-RBM15_(S+AE) fusion proteins, respectively, by introduction of theappropriate coding sequence into a host/vector expression system. Thereare numerous host/vectors systems available for the propagation ofnucleotide sequences and/or the production of expressed proteins. Theseinclude, but are not limited to, plasmid and viral vectors, andprokaryotic and eukaryotic host. One skilled in the art can readilyadapt any host/vector system which is capable of propagating orexpressing heterologous DNA to produce or express the sequences of thepresent invention. Of course, the RBM15-MKL1, MKL1-RBM15_(S) andMKL1-RBM15_(S+AE) fusion proteins or polypeptides derived therefrom mayalso be produced by other means known in the art such as, for example,chemical synthesis or in vitro transcription/translation.

[0064] Also provided by the present invention are an isolated RBM15-MKL1fusion protein (SEQ ID NO: 2), an isolated MKL1-RBM15_(S) fusion protein(SEQ ID NO: 4), and an isolated MKL1-RBM15_(S+AE) fusion protein (SEQ IDNO: 6), an isolated RBM15_(S) protein (SEQ ID NO: 8), an isolatedRBM15_(S+AE) protein (SEQ ID NO: 10), an isolated RBM15_(L) protein (SEQID NO: 12), and an isolated MKL1 protein (SEQ ID NO: 14), which areencoded by their cognate nucleotides, that is, by SEQ ID NO: 1, 3, 5, 7,9, 11, and 13, respectively. Synthetic oligopeptides derived from SEQ IDNO: 2, 4, 6, 8, 10, 12, and 14 are also provided in this embodiment ofthe invention.

[0065] An “isolated” or “purified” nucleic acid molecule or protein, orbiologically active portion thereof, is substantially free of othercellular material, or culture medium when produced by recombinanttechniques, or substantially free of chemical precursors or otherchemicals when chemically synthesized. Preferably, an “isolated” nucleicacid is free of sequences (preferably protein encoding sequences) thatnaturally flank the nucleic acid (i.e., sequences located at the 5′ and3′ ends of the nucleic acid) in the genomic DNA of the organism fromwhich the nucleic acid is derived. For purposes of the invention,“isolated” when used to refer to nucleic acid molecules excludesisolated chromosomes. For example, in various embodiments, the isolatednucleic acid molecules can contain less than about 5 kb, 4 kb, 3 kb, 2kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequences that naturally flankthe nucleic acid molecule in genomic DNA of the cell from which thenucleic acid is derived.

[0066] An isolated protein that is substantially free of cellularmaterial includes preparations of RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE), RBM15_(L), or MKL1 proteinhaving less than about 30%, 20%, 10%, or 5% (by dry weight) of non-likeprotein (also referred to herein as a “contaminating protein”). When theRBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S), RBM15_(S+AE),RBM15_(L), or MKL1 protein or biologically active portion thereof isrecombinantly produced, preferably, culture medium represents less thanabout 30%, 20%, 10%, or 5% of the volume of the protein preparation.When RBM15-MKL1, MKL1-RBM15_(S), MKL1-RBM15_(S+AE), RBM15_(S),RBM15_(S+AE), RBM15_(L), or MKL1 protein is produced by chemicalsynthesis, preferably the protein preparations have less than about 30%,20%, 10%, or 5% (by dry weight) of chemical precursors or non-likechemicals.

[0067] In Example 1, the present invention provides evidence that thenucleotide sequences containing the RBM15-MKL1 and MKL1-RBM15 fusiongenes are present in patients with t(1;22) AMKL. Based on thisobservation, the present invention provides methods of assaying for thepresence of nucleotide sequences containing the RBM15-MKL1 andMKL1-RBM15 fusions in a sample and thus provides an assay for thedetection of t(1;22) leukemias, as explained in Example 1. The methodsof the invention can involve any means known in the art for detectingthe presence of specific nucleotide sequences in a sample including, butnot limited to, nucleic acid hybridization and detection methods (e.g.,“Southerns,” “Northerns” and the like) fluorescence in situhybridization (FISH) and detection methods, or polymerase chain reaction(PCR) amplification and detection methods, particularly reversetranscriptase-polymerase chain reaction amplification (RT-PCR).

[0068] One example of the assay methods of the present invention whichare used to detect RBM15-MKL1 or MKL1-RBM15 fusion gene are based on thepreferential amplification of sequences within a sample which containthe nucleotide sequences encoding the RBM15-MKL1, MKL1-RBM15_(S) orMKL1-RBM15_(S+AE) fusion proteins. In one embodiment of the invention,RT-PCR is utilized to detect the t(1,22) rearrangement that isassociated with AMLK. The method involves the use of RT-PCR to detectthe presence of transcripts from the RBM15-MKL1 or MKL1-RBM15 fusiongenes. The method involves reverse transcription via reversetranscriptase of an RNA sample from a patient to produce cDNA. Forreverse transcription, an oligo-dT primer can be use, or alternatively,a primer designed to specifically anneal to RBM15-MKL1 mRNA,MKL1-RBM15_(S)mRNA, or MKL1-RBM15_(S+AE)mRNA can be employed to primecDNA synthesis. Such primers can be designed from the nucleotidesequences of the invention as set forth in SEQ ID NOS: 1, 3, and 5 usingmethods known to those of ordinary skill in the art. Then, PCRamplification of the cDNA can be performed utilizing primers designed toamplify at least a portion of the nucleotide sequences of to RBM15-MKL1,MKL1-RBM15_(S) or MKL1-RBM15_(S+AE). The amplified cDNA can be detectedby methods known in the art such as, for example, agarose gelelectrophoresis and ethidium bromide staining. The detection of thedesired cDNA corresponding to at least a portion of the RBM15-MKL1,MKL1-RBM15_(S) or MKL1-RBM15_(S+AE) indicates that the sample is from apatient with the t(1,22) rearrangement.

[0069] The methods of the invention involve the use of PCRamplification, particularly RT-PCR. Methods for PCR amplification areknown in the art. dligonucleotide primers can be designed for use in PCRreactions to amplify corresponding DNA sequences from genomic DNA orcDNA extracted from any organism of interest. Methods for PCRamplification and for designing PCR primers are generally known in theart and are disclosed in Sambrook et al. (1989) Molecular Cloning: ALaboratory Manual (2d ed., Cold Spring Harbor Laboratory Press,Plainview, N.Y.). See also Innis et al., eds. (1990) PCR Protocols: AGuide to Methods and Applications (Academic Press, New York); Innis andGelfand, eds. (1995) PCR Strategies (Academic Press, New York); andInnis and Gelfand, eds. (1999) PCR Methods Manual (Academic Press, NewYork). Other known methods of PCR that can be used in the methods of theinvention include, but are not limited to, methods using paired primers,nested primers, single specific primers, degenerate primers,gene-specific primers, mixed DNA/RNA primers, vector-specific primers,partially mismatched primers, and the like.

[0070] In addition to methods which rely on the amplification of atarget sequence, the present invention further provides methods foridentifying nucleic acids containing the RBM15-MKL1 fusion gene which donot require sequence amplification and are based on the known methods ofSouthern (DNA:DNA) and Northern (DNA:RNA) blot hybridizations, and FISHof chromosomal material, using probes derived from the nucleotidesequences of the invention. Additionally, other nucleotide sequences ofchromosomes 1 and 22 that are known to art and for which are disclosedherein to occur in the vicinity of the chromosomal breakpoint for thet(1;22)(p13;q13) chromosomal translocation event associated with AMKLcan be used in the methods of the invention. That is, nucleic acidprobes can be used that comprise nucleotide sequences in proximity tothe t(1;22)(p13;q13) chromosomal translocation event, or breakpoint. By“in proximity to” is intended within about 10 kilobases (kb) of thet(1;22) breakpoint. Such other nucleotide sequences include, but are notlimited to, RP11-260A24 (Accession no. AC025987), RP5-1042K10 (Accessionno. AL022238), RP11-313L7, RP5-1125M8 (Accession no. AL356387),RP4-665N5, RP4-743K1, RP11-50F6, RP3-377F16 (Accession no. Z93783),RP4-591N18 (Accession no. AL031594), RP1-229A8 (Accession no. Z86090),and RP4-735G18 (Accession no. AL096703). The clones not identified withAccession numbers are also available from the Roswell Park CancerInstitute (RPCI-BAC library).

[0071] In another embodiment of the invention, methods are provideddetecting the t(1,22) rearrangement involving FISH (fluorescence in situhybridization ) of human chromosomal material. For example, a probe thatis comprised of nucleotide sequences that span the breakpoint in eithera wild-type chromosome 1 or 22 can be used. Such a probe can hybridizeto both derivative chromosomes in the case of a t(1,22) rearrangement.Alternatively, two probes, each labeled with a different detectionreagent, can be utilized. The first probe is capable of hybridizing tosequences within the region of chromosomal band 1p13, and the secondprobe is capable of binding to the region of chromosomal band 22q13. Thetwo probes are also selected such that, in a t(1,22) rearrangement, bothprobes hybridize to the same derivative chromosome, whether it bechromosome 1 or 22. In such a case, a signal from each of the probes isobserved on the same chromosome.

[0072] The nucleic acid probes of the present invention include DNA aswell as RNA probes, such probes being generated using techniques knownin the art (Sambrook et al., eds., Molecular Cloning, Cold Spring HarborPress, Cold Spring Harbor, N.Y. (1989)). A skilled artisan can employsuch known techniques using the RBM15_(S), RBM15_(S+AE), RBM15_(L),MKL1, RBM15-MKL1, MKL1-RBM15_(S), and MKL1-RBM15_(S+AE) nucleotidesequences herein described, or fragments thereof, as probes.

[0073] For nucleic acid probes, examples of detection reagents include,but are not limited to radiolabeled probes, enzymatic labeled probes(horse radish peroxidase, alkaline phosphatase), and affinity labeledprobes (biotin, avidin, or steptavidin). For antibodies, examples ofdetection reagents include, but are not limited to, labeled secondaryantibodies, or in the alternative, if the primary antibody is labeled,the chromophoric, enzymatic, or antibody binding reagents which arecapable of reacting with the labeled antibody. One skilled in the artwill readily recognize that the antibodies and nucleic acid probesdescribed in the present invention can readily be incorporated into oneof the established kit formats which are well known in the art.

[0074] The samples used in the detection methods of the presentinvention include, but are not limited to, cells or tissues, protein,membrane, or nucleic acid extracts of the cells or tissues, andbiological fluids such as blood, serum, and plasma. The sample used inthe methods of the invention will vary based on the assay format, natureof the detection method, and the tissues, cells or extracts which areused as the sample. Methods for preparing protein extracts, membraneextracts or nucleic acid extracts of cells are well known in the art andcan be readily be adapted in order to obtain a sample which iscompatible with the method utilized (see, for example, K. Budelier etal., Chapter 2, “Preparation and Analysis of DNA,” M. E. Greenberg etal., Chapter 4, “Preparation and Analysis of RNA” and M. Moos et al.,Chapter 10, “Analysis of Proteins,” in Ausubel et al., Current Protocolsin Molecular Biology, Wiley Press, Boston, Mass. (1993)). One preferredtype of sample which can be utilized in the present invention is derivedfrom isolated lymphoma cells. Such cells can be used to prepare asuitable extract or can be used in procedures based on in situ analysis.

[0075] The present invention further provides antibodies specific toepitopes of the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusionproteins and methods of detecting the RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion proteins, or any combination thereof, that relyon the ability of these antibodies to selectively bind to specificportions of the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(s+AE)proteins that are unique to that fusion protein. Such antibodies do notbind preferentially to the RBM15 or MKL1proteins.

[0076] The present invention further provides methods of detecting thepresence of at least one of the RBM15-MKL1, MKL1-RBM15_(S) andMKL1-RBM15_(S+AE) fusion proteins. Antibodies can be prepared whichrecognize a fusion protein of the invention. Such antibodies can be usedto detect the presence of the fusion protein in samples from humancells. The methods of the invention involve the use of antibodies thatbind to at least one of the fusion proteins of the invention andantibody detection systems that are known to those of ordinary skill inthe art. Such methods find use in diagnosis and treatment of AMKL, forexample, to determine if particular cells or tissues express theRBM15-MKL1, MKL1-RBM15_(S), and/or the MKL1-RBM15_(S+AE) fusionproteins.

[0077] Conditions for incubating an antibody with a test sample varydepending on the format employed for the assay, the detection methodsemployed, the nature of the test sample, and the type and nature of theantibody used in the assay. One skilled in the art will recognize thatany one of the commonly available immunological assay formats (such asradioimmunoassays, enzyme-linked immunosorbent assays, diffusion basedouchterlony, or rocket inmunofluorescent assays) can readily be adaptedto employ the antibodies of the present invention. Examples of suchassays can be found in Chard, T., An Introduction to Radioimmunoassayand Related Techniques, Elsevier Science Publishers, Amsterdam, TheNetherlands (1986); Bullock, G. R. et al., Techniques inImmunocytochemistry, Academic Press, Orlando, Fla. Vol. 1 (1982), Vol. 2(1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of EnzymeImmunoassays: Laboratory Techniques in Biochemistry and MolecularBiology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).

[0078] In another embodiment of the immunoassays of the invention, theanti-RBM15-MKL1 antibody, the anti-MKL1-RBM15_(S) antibody, or theanti-MKL1-RBM15_(S+AE) antibody is immobilized on a solid support.Examples of such solid supports include, but are not limited to,plastics such as polycarbonate, complex carbohydrates such as agaroseand sepharose, and acrylic resins, such as polyacrylamide and latexbeads. Techniques for coupling antibodies to such solid supports arewell known in the art (see, for example, Weir, D. M. et al., Handbook ofExperimental Immunology, 4th Ed., Blackwell Scientific Publications,Oxford, England, Chapter 10 (1986)).

[0079] Additionally, one or more of the antibodies used in the abovedescribed methods can be detectably labeled prior to use. Antibodies canbe detectably labeled through the use of radioisotopes, affinity labels(such as biotin, avidin, etc.), enzymatic labels (such as horse radishperoxidase, alkaline phosphatase, etc.) fluorescent labels (such as FITCor rhodamine, etc.), paramagnetic atoms, etc. Procedures foraccomplishing such labeling are well-known in the art; see, for example,Stemberger, L. A. et al., J. Histochem. Cytochem. 18:315-333 (1970);Bayer, E. A. et al., Meth. Enzym. 62:308-315 (1979); Engrall, E. et al.,J. Immunol. 109:129-135 (1972); Goding, J. W., J. Immunol. Meth.13:215-226 (1976).

[0080] The present invention further includes methods for selectivelykilling cells expressing the RBM15-MKL1 fusion protein, theMKL1-RBM15_(S) fusion protein, and/or the MKL1-RBM15_(S+AE) fusionprotein by, for example, contacting a cell expressing the RBM15-MKL1,MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) fusion protein with a toxinderivatized antibody, wherein the antibody is capable of selectivelybinding to the fusion protein with only weak or no binding to non-fusionRBM15 or MKL1protein. An example of such an antibody is toxinderivatized antibodies which bind to the RBM15-MKL1 fusion proteinjunction. As used herein, an antibody is said to be “toxin-derivatized”when the antibody is covalently attached to a toxin moiety. Proceduresfor coupling such moieties to a molecule are well known in the art. Thebinding of a toxin derivatized antibody to a cell brings the toxinmoiety into close proximity to the cell and thereby promotes cell death.By providing such an antibody molecule to a mammal, the cell expressingthe fusion protein can be preferentially killed. Any suitable toxinmoiety may be employed; however, it is preferable to employ toxins suchas, for example, the ricin toxin, the cholera toxin, the diphtheriatoxin, radioisotopic toxins, or membrane-channel-forming toxins.

[0081] The antibodies or toxin-derivatized antibodies of the presentinvention may be administered to a mammal intravenously,intramuscularly, subcutaneously, enterally, topically or parenterally.When administering antibodies or peptides by injection, theadministration may be by continuous injections, or by single or multipleinjections.

[0082] The antibodies or toxin-derivatized antibodies of the presentinvention are intended to be provided to recipient mammal in a“pharmaceutically acceptable form” in an amount sufficient to“therapeutically effective.” An amount is said to be therapeuticallyeffective if the dosage, route of administration, etc. of the agent aresufficient to preferentially kill a portion of the cells expressing theRBM15-MKL1 or MKL1-RBM15 fusion protein. An antibody is said to be in a“pharmacologically acceptable form” if its administration can betolerated by a recipient patient. The antibodies of the presentinvention can be formulated according to known methods of preparingpharmaceutically useful compositions, whereby these materials, or theirfunctional derivatives, are combined with a pharmaceutically acceptablecarrier vehicle. Suitable vehicles and their formulation, inclusive ofother human proteins, e.g., human serum albumin, are described, forexample, in Remington's Pharmaceutical Sciences, 16th ed., Osol, A.,ed., Mack, Easton Pa. (1980). In order to form a pharmaceuticallyacceptable composition which is suitable for effective administration,such compositions will contain an effective amount of an antibody of thepresent invention together with a suitable amount of carrier. Inaddition to carriers, the antibodies of the present invention may besupplied in humanized form. Humanized antibodies may be produced, forexample by replacing an immunogenic portion of an antibody with acorresponding, but non-immunogenic portion (i.e., chimeric antibodies)(Robinson, R. R. et al., International Patent PublicationPCT/US86/02269; Akira, K. et al., European Patent Application 184,187;Taniguchi, M., European Patent Application 171,496; Morrison, S. L. etal., European Patent Application 173,494; Neuberger, M. S. et al., PCTApplication WO 86/01533; Cabilly, S. et al., European Patent Application125,023; Better, M. et al., Science 240:1041-1043 (1988); Liu, A. Y. etal., Proc. Natl. Acad. Sci. USA 84:3439-3443 (1987); Liu, A. Y. et al.,J. Immunol. 139:3521-3526 (1987); Sun, L. K. et al., Proc. Natl. Acad.Sci. USA 84:214-218 (1987); Nishimura, Y. et al., Cancer Res.47:999-1005 (1987); Wood, C. R. et al., Nature 314:446-449 (1985)); Shawel al., J. Natl. Cancer Inst. 80:1553-1559 (1988).

[0083] In providing a patient with an antibody or toxin-derivatizedantibody, the dosage of administered agent will vary depending upon suchfactors as the patient's age, weight, height, sex, general medicalcondition, previous medical history, etc. In general, it is desirable toprovide the recipient with a dosage of the antibody which is in therange of from about 1 pg/kg to 10 mg/kg (body weight of patient),although a lower or higher dosage may be administered.

[0084] The present invention also encompasses antisense nucleic acidmolecules, i.e., molecules that are complementary to a sense nucleicacid encoding a protein, e.g., complementary to the coding strand of adouble-stranded cDNA molecule, or complementary to an mRNA sequence.Accordingly, an antisense nucleic acid can hydrogen bond to a sensenucleic acid. The antisense nucleic acid can be complementary to anentire coding strand, or to only a portion thereof, e.g., all or part ofthe protein coding region (or open reading frame). An antisense nucleicacid molecule can be antisense to a noncoding region of the codingstrand of a nucleotide sequence encoding a protein of interest. Thenoncoding regions are the 5′ and 3′ sequences that flank the codingregion and are not translated into amino acids.

[0085] Given the coding-strand sequence encoding, for example, anRBM15-MKL1 fusion protein disclosed herein (e.g., SEQ ID NO: 1),antisense nucleic acids of the invention can be designed according tothe rules of Watson and Crick base pairing. The antisense nucleic acidmolecule can be complementary to the entire coding region of RBM15-MKL1mRNA, but more preferably is an oligonucleotide that is antisense toonly a portion of the coding or noncoding region of RBM15-MKL1 mRNA. Forexample, the antisense oligonucleotide can be complementary to theregion surrounding the translation start site of RBM15-MKL1 mRNA. Apreferred antisense oligonucleotide for selective hybridisation tofusion transcripts will include the region spanning the RBM15 portion ofthe fusion transcript and the MKL1 portion of the fusion transcript. Anantisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25,30, 35, 40, 45, or 50 nucleotides in length. An antisense nucleic acidof the invention can be constructed using chemical synthesis andenzymatic ligation procedures known in the art. Similarly, antisensenucleotide molecules can be prepared for the nucleotide sequencesencoding the MKL1-RBM15_(S), and MKL1-RBM15_(S+AE) fusion proteins (SEQID NOS: 3 and 5, respectively).

[0086] For example, an antisense nucleic acid (e.g., an antisenseoligonucleotide) can be chemically synthesized using naturally occurringnucleotides or variously modified nucleotides designed to increase thebiological stability of the molecules or to increase the physicalstability of the duplex formed between the antisense and sense nucleicacids, including, but not limited to, for example e.g., phosphorothioatederivatives and acridine substituted nucleotides. Alternatively, theantisense nucleic acid can be produced biologically using an expressionvector into which a nucleic acid has been subcloned in an antisenseorientation (i.e., RNA transcribed from the inserted nucleic acid willbe of an antisense orientation to a target nucleic acid of interest,described further in the following subsection).

[0087] When used therapeutically, the antisense nucleic acid moleculesof the invention are typically administered to a subject or generated insitu such that they hybridize with or bind to cellular mRNA and/orgenomic DNA encoding a protein of the invention to thereby inhibitexpression of the protein, e.g., by inhibiting transcription and/ortranslation. An example of a route of administration of antisensenucleic acid molecules of the invention includes direct injection at atissue site. Alternatively, antisense nucleic acid molecules can bemodified to target selected cells and then administered systemically.For example, antisense molecules can be linked to peptides or antibodiesto form a complex that specifically binds to receptors or antigensexpressed on a selected cell surface. The antisense nucleic acidmolecules can also be delivered to cells using the vectors describedherein. To achieve sufficient intracellular concentrations of theantisense molecules, vector constructs in which the antisense nucleicacid molecule is placed under the control of a strong pol II or pol IIIpromoter are preferred.

[0088] An antisense nucleic acid molecule of the invention can be anα-anomeric nucleic acid molecule. An cc-anomeric nucleic acid moleculeforms specific double-stranded hybrids with complementary RNA in which,contrary to the usual β-units, the strands run parallel to each other(Gaultier et al. (1987) Nucleic Acids Res. 15:6625-6641). The antisensenucleic acid molecule can also comprise a 2′-o-methylribonucleotide(Inoue et al. (1987) Nucleic Acids Res. 15:6131-6148) or a chimericRNA-DNA analogue (Inoue et al. (1987) FEBS Lett. 215:327-330).

[0089] In another embodiment of the present invention, methods areprovided for modulating the translation of at least one RNA selectedfrom the group consisting of those RNAs encoding the RBM15-MKL1,MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusion protein in the cell.Specifically, such methods comprise introducing into a cell a DNAsequence which is capable of transcribing RNA which is complimentary tothe mRNA encoding either the RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion protein. By introducing such a sequence into acell, antisense RNA will be produced that will hybridize to RBM15-MKL1,MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) mRNA and block the translation ofthe RBM15-MKL1 or MKL1-RBM15 fusion protein, respectively. Antisensecloning has been described elsewhere in more detail by Methis et al.,Blood 82:1395-1401 (1993); Stein et al., Science 261:1004-1012 (1993);Mirabella et al., Anti-Cancer Drug Design 6:647-661 (1991); Rosenberg etal., Nature 313:703-706 (1985); Preiss et al., Nature 313:27-32 (1985),Melton, Proc. Natl. Acad. Sci. USA 82:144-148 (1985) and Kim et al.,Cell 42:129-138 (1985). Transcription of the introduced DNA will resultin multiple copies of the antisense RNA being generated. By controllingthe level of transcription of antisense RNA, and the tissue specificityof expression via promoter selection or gene targeting of the antisenseexpression sequence, one skilled in the art can regulate the level oftranslation of the RBM15-MKL1, MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE)fusion proteins in specific cells within a patient. In a related method,one or more synthetic antisense oligonucleotides that are complementaryto the RBM15-MKL1, MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) codingsequences of the invention, optionally including chemical modificationsdesigned to stabilize the oligonucleotide or enhance its uptake intocells, are administered to cells of a patient by known methods (see, forexample, R. W. Wagner, Nature 372:333-335 (1994); J. Lisziewicz et al.,Proc. Natl. Acad. Sci. (USA) 90:3860-3864 (1993); S.Fitzpatrick-McElligott, Bio/Technology 10: 1036-1040 (1992); E. Uhlmannet al., Chemical Reviews 90:543-583 (1990); and B. Tseng et al., CancerGene Therapy 1:65-71 (1994)).

[0090] The invention also encompasses ribozymes, which are catalytic RNAmolecules with ribonuclease activity that are capable of cleaving asingle-stranded nucleic acid, such as an mRNA, to which they have acomplementary region. The level of expression of the RBM15-MKL1,MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) fusion proteins can also becontrolled through the use of ribozyme technology. Ribozymes (e.g.,hammerhead ribozymes (described in Haselhoff and Gerlach (1988) Nature334:585-591)) can be used to catalytically cleave RBM15-MKL1,MKL1-RBM1_(S), or MKL1-RBM15_(S+AE) mRNA transcripts to thereby inhibittranslation of RBM15-MKL1, MKL1-RBM15_(S), and MKL1-RBM15_(S+AE) mRNA,respectively. A ribozyme having specificity for an RBM15-MKL1-,MKL1-RBM15_(S), or MKL1-RBM₁₅ _(S+AE)-encoding nucleic acid can bedesigned based upon the nucleotide sequence of the corresponding cDNAdisclosed herein (e.g., SEQ ID NOS: 1, 3, and 5, respectively). See,e.g., Cech et al., U.S. Pat. No. 4,987,071; and Cech et al., U.S. Pat.No. 5,116,742. Alternatively, RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) mRNA can be used to select a catalytic RNA having aspecific ribonuclease activity from a pool of RNA molecules. See, e.g.,Bartel and Szostak (1993) Science 261:1411-1418.

[0091] The present invention further provides methods of generatingtransgenic animals and transformed cell lines which contain theRBM15-MKL1, MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) nucleotidesequences. Such animals and cell lines are useful as animal models forhuman t(1;22) leukemias. In general, methods of generating transgenicanimals and transformed cell lines are well known in the art (forexample, see Grosveld et al., Transgenic Animals, Academic Press Ltd.,San Diego, Calif. (1992)). Using the nucleotide sequences disclosedherein for the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) orcoding sequences for the RBM15-MKL1, MKL1-RBM15_(S), and/orMKL1-RBM15_(S+AE) fusion proteins, a skilled artisan can readilygenerate a transgenic animal and transformed cell lines which containsand expresses the RBM15-MKL1 fusion protein, the MKL1-RBM15_(S) fusionprotein, and/or MKL1-RBM15_(S+AE) fusion protein. Transgenic animals(such as mice and pigs) which express the RBM15-MKL1 fusion gene can beused as an animal model for human t(1;22) leukemia. Transgenic animalswhich express the RBM15-MKL1 fusion protein, the MKL1-RBM15_(S) fusionprotein, or MKL1-RBM15_(S+AE) fusion protein, or any combinationthereof, are useful for determining, at the molecular level, the rolesof the RBM15-MKL1, MKL1-RBM15_(S), and MKL1-RBM15_(S+AE) fusion proteinsin the development of acute megakaryoblastic leukemia. Such animalsserve as models for the development of alternative therapies for t(1;22)lymphoma.

[0092] Transformed eukaryotic cell lines that express on or more of thefusion of proteins of the invention can be used, for example, to screenfor agents that are useful for treating AMKL. Preferably such cell linesare mammalian cell lines. More preferably, such cell lines are humancell lines. Generally, desired agents are those that suppress oreliminate phenotypic changes that occur as a result of the expression ofone or more fusion proteins of the invention in the cell. Phenotypicchanges that occur as a result of the expression of one or more fusionproteins of the invention in the cell include, for example, the presenceof CD61 and absence of peroxidase and esterase (See, e.g. Bennett etal., Ann. Intern Med 103:460-462 (1985); Skinnider L. F. et al., ActaHaematologica 98 (1): 26 (1997); Avanzi et al., J. Cell Physiol. 145:458-464 (1990); Avanzi et al., Brit. J Haematol. 69: 359 (1988)).

[0093] Such desired agents may be further screened for selectivity bydetermining whether they suppress or eliminate phenotyic changes oractivities associated with expression of unfused RBM15 and/orMKL1proteins in cells that either express such unfused proteinsnaturally or are engineered to express such proteins. Selective agentsare those which suppress or eliminate phenotypes associated withexpression of the fusion protein but which do not suppress or eliminatethe phenotypes associated with the unfused RBM15 and MKL1 proteins.Typically, the agents are screened by administering the agent to thecell. It is recognized that it is preferable that a range of dosages ofa particular agent be administered to the cells to determine if theagent is useful for treating AMKL. Appropriate cell lines that can beused in this method include, but are not limited to, DAMI, MEG-01,M-07e, CMK, CHRF-288-11 and UT7 cells.In another embodiment of thepresent invention, methods are provided for identifying agents which arecapable of binding to the RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion proteins herein described. Such methodscomprise (a) contacting a candidate agent with RBM15-MKL1,MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusion protein, or fragmentthereof, and (b) determining whether the candidate agent binds to thefusion protein. Using this method, agents which can be used to modulatethe activity of the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE)fusion protein can be identified. Such methods can additionally comprisean additional step to select from the identified agents those which donot bind RBM15 or MKL1proteins. Such an additional step involvescontacting the agent with an RBM15 or MKL1 protein, or fragment thereofand determining whether the agent binds to the protein or fragment.

[0094] There are numerous variations of the above assays which can beused by a skilled artisan without the need for undue experimentation inorder to isolate agonists, antagonists, and ligands of the RBM15-MKL1,MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) fusion protein; see, forexample, Burch, R. M., in Medications Development. Drug Discovery,Databases, and Computer-Aided Drug Design, NIDA Research Monograph 134,NIH Publication No. 93-3638, Rapaka, R. S., and Hawks, R. L., eds., U.S.Dept. of Health and Human Services, Rockville, Md. (1993), pages 37-45.For example, an idiotypic antibody to RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion protein can be used to co-precipitate fusionprotein-bound agents in the purification and characterization of suchagents. Harlow, E., et al., Chapter 11 in Antibodies: A LaboratoryManual, Cold Spring Harbor Laboratories, Cold Spring harbor, N.Y.(1988), pages 421-470. Further, an anti-idiotypic antibody toRBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) can be used to designsynthetic RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) ligands.Ertl, H., et al., Vaccine 6:80-84 (1988); Wolff, M. E., in MedicationsDevelopment: Drug Discovery, Databases, and Computer-Aided Drug Design,NIDA Research Monograph 134, NIH Publication No. 93-3638, Rapaka, R. S.,and Hawks, R. L., eds., U.S. Dept. of Health and Human Services,Rockville, Md. (1993), pages 46-57. In addition, an anti-idiotypicantibody to the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusionproteins, the RBM15-MKL1, MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusionproteins, or a fragment thereof containing the active (ligand binding)site of the fusion protein, can be used to screen an expression libraryfor genes encoding proteins which bind the fusion protein.

[0095] Alternatively, cells expressing the RBM15-MKL1, MKL1-RBM15_(S),or MKL1-RBM15_(S+AE) fusion proteins on their surfaces can be used toscreen expression libraries or synthetic combinatorial oligopeptidelibraries. Cwirla, S. E., et al., Proc. Natl. Acad. Sci. (USA)87:6378-6382 (1990); Houghten, R. A., et al., Nature 354:84-86 (1991);Houghten, R. A., et al., in Medications Development: Drug Discovery,Databases, and Computer-Aided Drug Design, NIDA Research Monograph 134,NIH Publication No. 93-3638, Rapaka, R. S., and Hawks, R. L., eds., U.S.Dept. of Health and Human Services, Rockville, Md. (1993), pages 66-74.In particular, cells that have been genetically engineered to expressand display the RBM15-MKL1, MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE)fusion protein via the use of the nucleic sequences of the invention arepreferred in such methods, as host cell lines may be chosen which aredevoid of related receptors. Hartig, P. R., in Medications Development:Drug Discovery, Databases, and Computer-Aided Drug Design, NIDA ResearchMonograph 134, NIH Publication No. 93-3638, Rapaka, R. S., and Hawks, R.L., eds., U.S. Dept. of Health and Human Services, Rockville, Md.(1993), pages 58-65.

[0096] The agents screened in the above assay can be, but are notlimited to, small molecules, peptides, carbohydrates, or vitaminderivatives. The agents can be selected and screened at random orrationally selected or designed using protein modeling techniques. Forrandom screening, agents such as peptides or carbohydrates are selectedat random and are assayed for their ability to bind to the pseudogenepeptide. Alternatively, agents may be rationally selected or designed.As used herein, an agent is said to be “rationally selected or designed”when the agent is chosen based on the configuration of the pseudogenepeptide. For example, one skilled in the art can readily adapt currentlyavailable procedures to generate peptides capable of binding to aspecific peptide sequence in order to generate rationally designedantipeptide peptides, see, for example, Hurby et al., “Application ofSynthetic Peptides: Antisense Peptides,” in Synthetic Peptides: A User'sGuide, W. H. Freeman, New York (1992), pp. 289-307; and Kaspczak et al.,Biochemistry 28:9230-2938 (1989).

[0097] Using the above procedures, the present invention provides agentscapable of binding to the the RBM15-MKL1, MKL1-RBM15_(S), and/orMKL1-RBM15_(S+AE) fusion proteins, produced by a method comprising thesteps of (a) contacting said agent with the the RBM15-MKL1,MKL1-RBM15_(S), and/or MKL1-RBM15_(S+AE) fusion protein, or a fragmentthereof, and (b) determining whether said agent binds to the RBM15-MKL1,MKL1-RBM15_(S), or MKL1-RBM15_(S+AE) fusion protein. Additional step(s)to determine whether such binding is selective for the fusion proteinrelative to the corresponding unfused RBM15 and MKL1 proteins may alsobe employed.

[0098] The materials used in the above assay methods (both nucleic acidand protein based) are ideally suited for the preparation of a kit. Forexample, for amplification based detection systems, the inventionprovides a compartmentalized kit to receive in close confinement, one ormore containers which comprises (a) a first container comprising one ormore of the amplification primers of the present invention, and (b) oneor more other containers comprising one or more of the following: asample reservoir, amplification reagents, wash reagents, and detectionreagents.

[0099] For antibody based detection systems, the present inventionprovides a compartmentalized kit to receive in close confinement, one ormore containers which comprises (a) a first container comprising anantibody capable of binding to the RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion protein and (b) one or more other containerscomprising one or more of the following: wash reagents and reagentscapable of detecting the presence of bound antibodies from the first andthe second containers.

[0100] The invention further provides a kit compartmentalized to receivein close confinement one or more containers which comprises (a) a firstcontainer comprising an antibody capable of binding to an epitope whichis present in the fusion junction of the RBM15-MKL1, MKL1-RBM15_(S), orMKL1-RBM15_(S+AE) fusion protein and which is not present in either ofthe two non-fusion proteins; and (b) one or more other containerscomprising one or more of the following: wash reagents and reagentscapable of detecting the presence of bound antibodies from the firstcontainer.

[0101] In detail, a compartmentalized kit includes any kit in whichreagents are contained in separate containers. Such containers includesmall glass containers, plastic containers or strips of plastic orpaper. Such containers allow one to efficiently transfer reagents fromone compartment to another compartment such that the samples andreagents are not cross-contaminated, and the agents or solutions of eachcontainer can be added in a quantitative fashion from one compartment toanother. Such containers may include a container which will accept thetest sample, a container which contains the antibodies or probes used inthe assay, containers which contain wash reagents (such as phosphatebuffered saline, Tris-buffers, etc.), and containers which contain thereagents used to detect the bound antibody or the hybridized probe. Anydetection reagents known in the art can be used including, but notlimited to those described supra.

[0102] The following examples are offered by way of illustration and notby way of limitation.

EXPERIMENTAL EXAMPLE 1 Fusion of RNA Recognition Motif-Encoding Gene,RBM15, to the SAP DNA-Binding Domain Gene, MegaKaryoblastic Leukemia-1(MKL1), in Acute Megakaryoblastic Leukemias with t(1;22)(p13;q13)Summary

[0103] Acute megakaryoblastic leukemia (AMKL) in young children isalmost invariably caused by leukemic blasts harboring t(1;22)(p13 ;q13)(Carroll, A. et al., Blood 78:748-752 (1991); Lion, T. et al., Blood79:3325-3330 (1992); Bernstein, J. et al., Leukemia 14:216-218 (2000)).Despite its remarkable disease specificity and a lack of knowledge ofAMKL pathogenesis (Cripe, L. D. & Hromas, R., Semin. Hematol. 35:200-209(1998)), t(1;22) has yet to be characterized molecularly. Disclosedherein is the identification of the reciprocal fusion transcriptsderived from two novel genes, RNA-binding motif protein-15 (RBM15) atchromosome p13 and Megakaryoblastic Leukemia-1 (MKL1) at 22q13, as theconsequence of t(1;22). RBM15, detected in three isoforms—RBM15_(L),RBM15_(S), and RBM15_(S+AE)—contains three RNA recognition motifs (RRM)(Burd, C. G. & Dreyfuss, G., Science 265:615-621 (1994)) and a Spenparalog and ortholog C-terminal (SPOC) domain (Wiellette, E. L. et al.,Development 126:5373-5385 (1999)), thus showing significant homology tospen, a homeotic Drosophila gene capable of enhancing Ras/MAP kinasesignaling (Wiellette, E. L. et al., Development 126:5373-5385 (1999);Rebay, I. et al., Genetics 154:695-712 (2000); Chen, F. & Rebay, I.,Curr. Biol. 10:943-946 (2000); Kuang, B. et al., Development127:1517-1529 (2000)). MKL1 contains a SAP (SAF-A/B, Acinus and PIAS)DNA-binding motif (Aravind, L. & Koonin, E. V., Trends Biochem. Sci.25:112-114 (2000)) that in homologous proteins such as SAF-B functionsto recruit domains involved in chromatin remodeling, transcriptionalcontrol, and pre-mRNA processing to the matrix attachment regions (MAR)of transcriptionally active chromatin, effectively couplingtranscription and splicing (Naylor, O. et al., Nucleic Acids Res.26:3542-3549 (1998)). Although both reciprocal fusion transcripts areexpressed in AMKL, RBM15-MKL1, from the der(22) chromosome, encodes allputative functional motifs of each gene and is the candidate oncogene oft(1;22), through a mechanism that may involve deregulation of RNAprocessing and/or Hox and Ras/MAP kinase signaling.

Description

[0104] Cloning of chromosomal translocations has led to identificationof pathogenically relevant oncogenic fusion transcripts and proteins inspecific subsets of acute nonlymphocytic leukemia (ANLL), such aspromyelocytic leukemia-retinoic acid receptor alpha fusion gene(PML-RARα) in acute promyelocytic leukemia (FAB-M3 subtype), acutemyeloid leukemia 1-eight twenty one fusion gene (AML1-ETO) in ANLL withmaturation (FAB-M2), and various mixed lineage leukemia (MLL) genefusions in acute myelomonocytic and monocytic leukemias (FAB-M4 and -M5)(Melnick, A. & Licht, J. D., Blood 93:3167-3215 (1999); Downing, J. R.,Br. J. Haematol. 106:296-308 (1999); Rowley, J. D., Semin. Hematol.36:59-72 (1999); Look, A. T., Science 278:1059-1064 (1997); Faretta, M.,Di Croce, L. & Pelicci, P. G., Sem. Hematol. 38:42-53 (2001)). Despitethese significant advances, little is known about the genetic mechanismsunderlying acute leukemias of the megakaryoblastic (platelet precursor)lineage (AMKL, FAB-M7) (Cripe, L. D, infra). Almost invariably, AMKL innon-Down syndrome infants and young children harbor thet(1;22)(p13;q13), in most cases as the sole cytogeneticabnormality(Carroll, A. et al; Lion, T. et al., and Bernstein, J. etal., infra). Phenotypically, AMKL presents de novo (i.e., without aso-called preleukemic stage), with a large leukemia cell mass, andfrequent fibrosis of bone marrow and other organs. Progression isusually rapid despite therapy, with a median overall patient survival ofonly 8 months.

[0105] To clone t(1;22), a fluorescence in situ hybridization(FISH)-based positional cloning strategy was used to define the 1p13 and22q13 breakpoints . Bacterial artificial chromosome (BAC) clones mappingto each chromosomal band were selected using the public and Celera humansequence databases and used pairwise in a series of two-color, two-probeFISH analyses of metaphase chromosomes from t(1;22)-containing leukemiablasts to identify closely flanking clones. With this strategy, a singlechromosome 22 BAC clone, RP5-1042K10, was found that hybridized to boththe der(1) and der(22) chromosomes formed by the reciprocal balancedt(1;22) and thus contained the altered 22q13 gene locus. Additional FISHexperiments using DNA subfragments of RP5-1042K10 prepared bylong-distance PCR (LD-PCR) methods allowed refinement of the chromosome22 breakpoint to within the most telomeric gene in this clone, andrevealed that all cases with t(1;22) possessed genomic breakpoints in asingle 28-kb intron of this gene. The same approach demonstrated thebreakpoint on chromosome 1p13 to be encompassed by BAC clone RP11-260A24. After complete annotation of the sequence by combiningentries in the public and Celera databases, the 1p13 breakpoint wassublocalized to within an 6-kb genomic interval (extending fromnucleotide 1,798,871 to 1,804,858 in Celera scaffold GA_x2HTBL4WN8M)using RP11-260A24 LD-PCR subfragments in metaphase FISH. FISH analyseswith probes closely flanking the identified breakpoint regions onchromosome 1 and chromosome 22 confirmed the results of our ‘splitsignal’ analysis, highlighting both the der(1) and der(22) chromosomes.The breakpoint-spanning clones from each chromosome identified by FISHanalysis are RP11-260A24 (chromosome 1) and RP5-1042K10 (chromosome 22).An additional 22q13 BAC, clone RP11-313L7, that spanned the breakpointwas subsequently also identified, the end sequences (accession nos.AQ506839 (Sp6) and AQ537696 (T7)) of which revealed a 65,460 bp overlapwith RP5-1042K10 and a 76,650 bp overlap with an additional chromosome22 clone for this region designated RP4-591N18.

[0106] Database searches using the exon sequences flanking thebreakpoint-containing intron on chromosome 22 identified an anonymoushuman brain cDNA library clone (accession no. AB037859). This 3,907-bpcDNA encoded a 2,793-nucleotide ORF, with the putative ATG initiatorcodon in a context (ATCatgC) adequate to support translationalinitiation (Kozak, M., Mammalian Genome 7:563-574 (1996)). Because RNAblot hybridizations using this clone revealed an approximately 4.5-kbtranscript expressed ubiquitously in normal human tissues, additional 5′untranslated sequence was obtained by RACE, resulting in a complete cDNAof 4,447 bp (SEQ ID NO: 13). To denote its involvement in AMKL, thisgene was named MKL1 (eg4aaryoblastic Leukemia-1, official HUGONomenclature Committee designation). Motif searches of the deduced931-amino acid (aa) MKL1 protein (predicted mass, 98.9 kDa; SEQ ID NO:14) identified a bipartite nuclear localization signal (BP-NLS)(residues 14-31 of SEQ ID NO: 14; RRSLERARTEDYLKRKIR), a single SAPDNA-binding motif (Aravind, L. & Koonin, E. V., Trends Biochem. Sci. 25,112-114 (2000)) (residues 347-381 of SEQ ID NO: 14), a coiled-coilregion (residues 521-563 of SEQ ID NO: 14) that likely mediates proteinoligomerization (Lupas, A., Trends Biochem. Sci. 21, 375-382 (1996)),and a long C-terminal proline-rich segment (residues 564-811 of SEQ IDNO: 14) similar to proline-rich regions shown to act as transcriptionalactivators (Mitchell, P. J. & Tjian, R., Science 245, 372-378 (1989)).In addition, a short glutamine-rich segment (residues 264-286 of SEQ IDNO: 14; QQQQLFLQLQILNQQQQQHHNYQ) was found that is highly reminiscent ofthe more extensive glutamine-rich regions of the MLL acuteleukemia-associated transcription factor family, as well as a number ofother proteins involved in transcriptional control (Prasad, R. et al.,Oncogene 15, 549-560 (1997)). Of note, MKL1 showed significantcross-species homology to the product of a Drosophila gene ofundetermined function, CG12188 (accession no. AAF47681), exhibiting 41%identity (57% similarity) over the initial 161 amino acids of MKL1 and63% identity (76% similarity) in the SAP domains of the two proteins.

[0107] The MKL1 SAP domain shares sequence similarities with SAP domainsfrom (a) THO1—yeast protein Tho1p, which regulates transcriptionalelongation by RNA polymerase II; (b) E1B-55kDa, a transformingadenovirus protein that binds and inhibits p53, and mediatesnucleocytoplasmic transport of adenoviral and cellular mRNAs; (c) PIAS1(protein inhibitor of activated StatI) which binds and inhibits Stat1,coactivates transcription by various steroid receptors, regulates RNAhelicase II function, and has also been reported to bind wt and mutantp53; (d) SAF-B (scaffold attachment factor B), a RRM-containing proteinthat binds both RNA polymerase II and a subset of serine-/arginine-richRNA splicing factors; and (e) ACINUS (apoptotic chromatin condensationinducer in the nucleus) which mediates chromatin condensation duringprogrammed cell death (reviewed in Aravind, L. & Koonin, E. V., TrendsBiochem. Sci. 25, 112-114 (2000)).

[0108] With the hypothesis that t(1;22) generates an oncogenic fusionanalogous to breakpoint cluster region-Abelson tyrosine kinase fusiongene (BCR-ABL) or the Mixed-lineage leukemia (MLL)fusion genes inleukemias, 5′ RACE was performed with total RNA from our leukemiapatient samples using MKL1 oligonucleotide primers to identify the 1p13fusion partner. The obtained sequences corresponded to two anonymous,partially overlapping cDNA clones (accession nos. AK025596, AK022541)from chromosome 1 that were also contained within the approximately 6-kbgenomic interval in BAC RP11-260A24 previously demonstrated by FISH tospan the p13 breakpoint. Due to the presence of three RNA recognitionmotifs (RRM) encoded by these sequences, the corresponding gene wasnamed RBM15 (RNA-binding motif protein-15, official HUGO NomenclatureCommittee name). Using 5′ RACE and human placenta cDNA libraryscreening, the complete RBM15 coding sequence was obtained byidentification of an ATG initiator codon 132 nucleotides (nts) upstreamof the previously deposited sequences and preceded 30 nts by an in-frameTGA stop. Sequencing of RT-PCR products obtained with RBM15-specificprimers from normal leukocyte mRNA demonstrated three transcripts thatshare an identical 2,863-bp 5′ coding sequence, differing only in theirextreme 3′ coding portions due to alternative exon usage (FIG. 1)

[0109] RBM15 contains three RRM motifs and a BP-NLS in its C-terminus,and is highly homologous to Drosophila gene product GH11110 (accessionno. AF 145664) (40% identity, 56% similarity with the RRM-containingregion of RBM15 from residues 170-529; 39% identity, 54% similarity withthe RBM15 C-terminus from residues 714-954; percent identity determinedusing BLAST 2.1.3 (Altschul, S. F. et al., Nucleic Acids Res. 25: 3389(1997)) with default parameters selected). These regions of RBM15 andDrosophila GH11110 (also previously referred to as D. melanogaster Shortspen-like protein-2, DmSSLP2) (Wiellette, E. L. et al., Development 126,5373-5385 (1999)) are closely related to Drosophila spen (split ends)—anRRM protein that modulates Hox homeotic function (e.g., cooperating withAntennapedia to suppress head-like development in the thoracic region),and regulates neuronal cell fate and axon extension by enhancing Ras/MAPkinase signaling (Wiellette, E. L. et al., Development 126, 5373-5385(1999); Rebay, I. et al., Genetics 154, 695-712 (2000); Chen, F. &Rebay, I., Curr. Biol. 10, 943-946 (2000); Kuang, B. et al., Development127, 1517-1529 (2000)). Thus, the RBM15 C-terminus also contains aso-called SPOC (Spen paralog and ortholog C-termninal) domain, a 165-aaconserved motif of undetermined function found in Spen and Spen-likeproteins (Wiellette, E. L. et al., Development 126, 5373-5385 (1999))including the mammalian spen ortholog, MINT (Msx2-interacting nucleartarget), which binds homeoprotein Msx2 (Hox 8) and coregulatesosteoblast gene expression during craniofacial development (Newberry, E.P. et al., Biochemistry 38, 10678-10690 (1999)).

[0110] In t(1;22)-positive AMKL blasts, RT-PCR demonstrated expressionof both reciprocal fusion transcripts, RBM15-MKL1 and MKL1-RBM15. RT-PCRreactions using RBM15 sense (RBM15(S)-2746F) and MKL1 antisense(MKL1-204R) primers amplify a single 268-bp RBM15-MKL1product in allpatients (nucleotides 2866 to 3133 of SEQ ID NO: 1). Using MKL1 sense(MKL1-F) and RBM15 antisense (RBM15(S)-2930R, corresponding to sequencesof the 3′ most exon unique to RBM15_(S) and RBM15_(S+AE)) primers, tworeciprocal MKL1-RBM15 fusion transcripts are detected, one (251 bp;nucleotides 411 to 661 of SEQ ID NO: 3) containing the 3′ sequences fromRBM15_(S) and the other (362 bp; nucleotides 411 to 772 of SEQ ID NO: 5)with 3′ sequences found in RBM15_(S+AE) . No MKL1-RBM15 RT-PCR productswere obtained using MKL1-F and an antisense primer specific forRBM15_(L) (RBM15(L)-1636R) in any patients examined.

[0111] The predicted RBM15-MKL1 chimeric protein encoded on the der(22)contains all putative functional motifs of each normal protein (FIG. 1).Frequent duplication of der(1) in t(1;22)-containing blasts has led tospeculation that this abnormal chromosome likely encodes the oncogenicAMKL fusion protein(s) (Carroll, A. et al.; Lion, T. et al., Bernstein,J. et al., infra.); however, a functional role for MKL1-RBM15_(S) andMKL1-RBM15_(S+AE) in leukemogenesis is unclear given they encodepredicted products of only 17 and 25 aa, respectively.

[0112] The SAP motif mediates DNA binding of proteins to the AT-richmatrix attachment regions (MAR) associated with transcriptionally activechromatin (Aravind, L. & Koonin, E. V., Trends Biochem. Sci. 25, 112-114(2000)). SAP proteins include SAF-B (Chen, F. & Rebay, I., Curr. Biol.10, 943-946 (2000)), involved in RNA processing; Acinus (Sahara, S. etal., Nature 401, 168-173 (1999)), which induces chromatin condensation;and PIAS proteins (Valdez, B. C. et al., Biochem. Biophys. Res. Commun.234, 335-340 (1997); Chung, C. D. et al., Science 278, 1803-1805 (1997);Kotaja, N. et al., Mol. Endocrinol. 14, 1986-2000 (2000)) that bind RNAhelicase II, inhibit STAT signal transduction, and modulate steroidreceptor-dependent transcription. Thus, the SAP targets a diverse set offunctional domains to MAR sequences, coupling transcription andsplicing. In addition to modulating homeotic protein functions (and inthe case of spen, enhancing Ras/MAP kinase signals), Spen familyproteins like MINT can bind specific DNA sequences via their RRMdomains, an RRM function seen also in other transcriptional regulatorssuch as sea urchin SSAP (Stage-specific activator protein) (DeAngelo, D.J. et al., Mol. Cell. Biol. 15, 1254-1264 (1995); DeFalco, J. & Childs,G., Proc. Natl. Acad Sci. 93, 5802-5807 (1996)) and hTAFII68, anRNA/ssDNA-binding protein homologous to pro-oncoproteins TLS/FUS and EWS(Bertolotti, A. et al., EMBO J 15, 5022-5031 (1996); Bertolotti, A. etal., Oncogene 18, 8000-8010 (1999)). In RBM15-MKL1, the MKL1 SAP motifwould be expected to relocalize the RRM domains of RBM15 aberrantly tosites of transcriptionally active chromatin, targeting genes criticalfor the normal proliferation or differentiation of megakaryoblasts.

Methods

[0113] Clinical Cases

[0114] Leukemia specimens with histopathological and immunophenotypicfeatures typical of AMKL were studied from five infants and youngchildren. All cases contained t(1;22)(p13;q13) with the exception ofpatient 2, whose blasts possessed a complex t(1;6;22)(p13;p12;q13). Allfive specimens were shown to contain rearrangement of RBM15 and MKL1 byRT-PCR and/or FISH analysis.

[0115] Fluorescence In Situ Hybridization (FISH)

[0116] DNA was labeled by nick translation with digoxigenin-11-dUTPand/or biotin-16-dUTP (Roche Molecular Biochemicals). Labeled probeswere combined with sheared human DNA and hybridized to fixed interphasenuclei and metaphase cells in 50% formamide, 10% dextran sulfate and2×SSC at 37° C. and subsequently washed in a 50% formamide, 2×SSCsolution at 37° C. Hybridization signals were detected withfluorescein-labeled anti-digoxigenin (Ventana Medical Systems) fordigoxigenin-labeled probes and Texas red-avidin for biotinylated probes.Chromosomes and nuclei were stained with 4,6-diamidino-2-phenylindole(DAPI) prior to analysis.

[0117] Rapid Amplification of cDNA Ends (RACE) and DNA Sequencing

[0118] Total RNA was extracted from t(1;22)-positive frozen AMKLspecimens using RNA STAT-60 (Tel-Test, Inc.). Approximately 0.2 μg RNAwas used for 5′ RACE experiments that identified RBM15 as the chromosome1p13 partner gene of MKL1. Reverse transcription was performed withprimer MKL1-294R (SEQ ID NO: 15). After purification and tailing of thecDNA, PCR was performed with an oligo-dT anchor primer and MKL1 reverseprimer MKL1-73R (SEQ ID NO: 16), using temperature cycling conditionsrecommended by the manufacturer (Roche Molecular Biochemicals) and aThermal Cycle Model 2400 (Perkin-Elmer Cetus). Twenty microliters of PCRproduct were separated on a 2% agarose gel, and specific bands werepurified (Qiaquick gel extraction kit, Qiagen, Inc.) and sequenced usingMKL1 reverse primer MKL1-59R (SEQ ID NO: 17).

[0119] RT-PCR of RBM15-MKL1 and MKL1-RBM15 Fusion Transcripts

[0120] For RBM15-MKL1 detection, 1 μg of total RNA was reversetranscribed using primer MKL1-294R (SEQ ID NO: 15). PCR was performedusing primers RBM15(S)-2746F (SEQ ID NO: 18) and MKL1-204R (SEQ ID NO:19) and 35 cycles (94° C. for 15 s, 60° C. for 30 s, 72° C. for 30 s).Primer pair MKL1-F (SEQ ID NO: 20) and MKL1-204R, designed to amplify aportion of the ubiquitously expressed MKL1, were included in controlexperiments to verify RNA quality and RT-PCR technique. PCR productswere gel purified, then cycle sequenced using primers RBM15(S)-2746F andMKL1-204R. For detection of MKL1-RBM15 fusion transcripts, reversetranscription was done using an oligo-dT primer and PCR performed withprimers MKL1-F (SEQ ID NO: 20) and RBM15(S)-2930R (SEQ ID NO: 21) (toidentify MKL1-RBM15_(S) and MKL1-RBM15_(S+AE)) or MKL1-F (SEQ ID NO: 20)and RBM15(L)-1636R (SEQ ID NO: 22) (to detect MKL1-RBM15L).Amplification of normal RBM15 sequences for quality control wasperformed with primer pairs RBM15(S)-2746F (SEQ ID NO: 18) andRBM15(S)-2930R (SEQ ID NO: 21) or RBM15(L)-1636R (SEQ ID NO: 22).

[0121] Northern Blot Analysis

[0122] Normal human peripheral blood leukocyte total RNA was extracted(RNEasy kit, Qiagen), then treated with RNase-free DNase for 15 m atroom temp. This RNA was reverse transcribed and used as the template inPCR amplifications (35 cycles: 94° C. for 10 s, 62° C. for 10 s, 68° C.for 1 m) to generate cDNA fragments corresponding to probes a-d (FIG.1). The following PCR primer pairs were used: RBM15 probe a (433 bp),RBM15-1118F (SEQ ID NO: 23) and RBM15-1551R (SEQ ID NO: 24); RBM15 probeb (318 bp), RBM15-2831F (SEQ ID NO: 25) and RBM15-3149R (SEQ ID NO: 26);RBM15 probe c (388 bp), RBM15-1616F (SEQ ID NO: 27) and RBM15-2004R (SEQID NO: 28); MKL1 probe d (449 bp), MKL1-155F (SEQ ID NO: 29) andMKL1-294R (SEQ ID NO: 15). Multiple tissue Northern blots (Clontech)containing approximately 2 μg of poly(A)+RNA prepared from normal humantissues were hybridized at 68° C. for 2 h in ExpressHyb buffer(Clontech) using these RBM15 and MKL1 cDNA probes or a β-actin probesupplied by the manufacturer. Filters were autoradiographed at −80° C.with one intensifying screen for 3 d (probes a, c and d), 1 d (probe b)or 1 h (for β-actin).

[0123] All publications and patent applications mentioned in thespecification are indicative of the level of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

[0124] Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the following embodiments.

1 29 1 6836 DNA Homo sapiens CDS (84)..(5732) 1 ggcgccttcc tggagcgcggggagatgtaa agatagacaa ataattttcc caatgagact 60 gtagaagaga gagcaattgg ccaatg agg act gcg ggg cgg gac cct gtg ccg 113 Met Arg Thr Ala Gly Arg AspPro Val Pro 1 5 10 cgg cgg agt cca aga tgg cgg cgt gcg gtt ccg ctg tgtgaa acg agc 161 Arg Arg Ser Pro Arg Trp Arg Arg Ala Val Pro Leu Cys GluThr Ser 15 20 25 gcg ggg cgg cgg gtt act cag ctc cgc gga gac gac ctc cgacga ccc 209 Ala Gly Arg Arg Val Thr Gln Leu Arg Gly Asp Asp Leu Arg ArgPro 30 35 40 gca aca atg aag gga aaa gag cgc tcg cca gtg aag gcc aaa cgctcc 257 Ala Thr Met Lys Gly Lys Glu Arg Ser Pro Val Lys Ala Lys Arg Ser45 50 55 cgt ggt ggt gag gac tcg act tcc cgc ggt gag cgg agc aag aag tta305 Arg Gly Gly Glu Asp Ser Thr Ser Arg Gly Glu Arg Ser Lys Lys Leu 6065 70 ggg ggc tct ggt ggc agc aat ggg agc agc agc gga aag acc gat agc353 Gly Gly Ser Gly Gly Ser Asn Gly Ser Ser Ser Gly Lys Thr Asp Ser 7580 85 90 ggc ggt ggg tcg cgg cgg agt ctc ctc ctg gac aag tcc agc agt cga401 Gly Gly Gly Ser Arg Arg Ser Leu Leu Leu Asp Lys Ser Ser Ser Arg 95100 105 ggt ggc agc cgc gag tat gat acc ggt ggg ggc agc tcc agt agc cgc449 Gly Gly Ser Arg Glu Tyr Asp Thr Gly Gly Gly Ser Ser Ser Ser Arg 110115 120 ttg cat agt tat agc tcc ccg agc acc aaa aat tct tcg ggc ggg ggc497 Leu His Ser Tyr Ser Ser Pro Ser Thr Lys Asn Ser Ser Gly Gly Gly 125130 135 gag tcg cgc agc agc tcc cgg ggt gga ggc ggg gag tca cgt tcc tct545 Glu Ser Arg Ser Ser Ser Arg Gly Gly Gly Gly Glu Ser Arg Ser Ser 140145 150 ggg gcc gcc tcc tca gct ccc ggc ggc ggg gac ggc gcg gaa tac aag593 Gly Ala Ala Ser Ser Ala Pro Gly Gly Gly Asp Gly Ala Glu Tyr Lys 155160 165 170 act ctg aag ata agc gag ttg ggg tcc cag ctt agt gac gaa gcggtg 641 Thr Leu Lys Ile Ser Glu Leu Gly Ser Gln Leu Ser Asp Glu Ala Val175 180 185 gag gac ggc ctg ttt cat gag ttc aaa cgc ttc ggt gat gta agtgtg 689 Glu Asp Gly Leu Phe His Glu Phe Lys Arg Phe Gly Asp Val Ser Val190 195 200 aaa atc agt cat ctg tcg ggt tct ggc agc ggg gat gag cgg gtagcc 737 Lys Ile Ser His Leu Ser Gly Ser Gly Ser Gly Asp Glu Arg Val Ala205 210 215 ttt gtg aac ttc cgg cgg cca gag gac gcg cgg gcg gcc aag catgcc 785 Phe Val Asn Phe Arg Arg Pro Glu Asp Ala Arg Ala Ala Lys His Ala220 225 230 aga ggc cgc ctg gtg ctc tat gac cgg cct ctg aag ata gaa gctgtg 833 Arg Gly Arg Leu Val Leu Tyr Asp Arg Pro Leu Lys Ile Glu Ala Val235 240 245 250 tat gtg agc cgg cgc cgc agc cgc tcc cct tta gac aaa gatact tat 881 Tyr Val Ser Arg Arg Arg Ser Arg Ser Pro Leu Asp Lys Asp ThrTyr 255 260 265 cct cca tca gcc agt gtg gtc ggg gcc tct gta ggt ggt caccgg cac 929 Pro Pro Ser Ala Ser Val Val Gly Ala Ser Val Gly Gly His ArgHis 270 275 280 ccc cct gga ggt ggt gga ggc cag aga tca ctt tcc cct ggtggc gct 977 Pro Pro Gly Gly Gly Gly Gly Gln Arg Ser Leu Ser Pro Gly GlyAla 285 290 295 gct ttg gga tac aga gac tac cgg ctg cag cag ttg gct cttggc cgc 1025 Ala Leu Gly Tyr Arg Asp Tyr Arg Leu Gln Gln Leu Ala Leu GlyArg 300 305 310 ctg ccc cct cca cct ccg cca cca ttg cct cga gac ctg gagaga gaa 1073 Leu Pro Pro Pro Pro Pro Pro Pro Leu Pro Arg Asp Leu Glu ArgGlu 315 320 325 330 aga gac tac ccg ttc tat gag aga gtg cgc cct gca tacagt ctt gag 1121 Arg Asp Tyr Pro Phe Tyr Glu Arg Val Arg Pro Ala Tyr SerLeu Glu 335 340 345 cca agg gtg gga gct gga gca ggt gct gct cct ttc agagaa gtg gat 1169 Pro Arg Val Gly Ala Gly Ala Gly Ala Ala Pro Phe Arg GluVal Asp 350 355 360 gag att tca ccc gag gat gat cag cga gct aac cgg acgctc ttc ttg 1217 Glu Ile Ser Pro Glu Asp Asp Gln Arg Ala Asn Arg Thr LeuPhe Leu 365 370 375 ggc aac cta gac atc act gta acg gag agt gat tta agaagg gcg ttt 1265 Gly Asn Leu Asp Ile Thr Val Thr Glu Ser Asp Leu Arg ArgAla Phe 380 385 390 gat cgc ttt gga gtc atc aca gaa gta gat atc aag aggcct tct cgc 1313 Asp Arg Phe Gly Val Ile Thr Glu Val Asp Ile Lys Arg ProSer Arg 395 400 405 410 ggc cag act agt act tac ggc ttt ctc aaa ttt gagaac tta gat atg 1361 Gly Gln Thr Ser Thr Tyr Gly Phe Leu Lys Phe Glu AsnLeu Asp Met 415 420 425 tct cac cgg gcc aaa tta gca atg tct ggc aaa attata att cgg aat 1409 Ser His Arg Ala Lys Leu Ala Met Ser Gly Lys Ile IleIle Arg Asn 430 435 440 cct atc aaa att ggt tat ggt aaa gct aca ccc accacc cgc ctc tgg 1457 Pro Ile Lys Ile Gly Tyr Gly Lys Ala Thr Pro Thr ThrArg Leu Trp 445 450 455 gtg gga ggc ctg gga cct tgg gtt cct ctt gct gccctg gca cga gaa 1505 Val Gly Gly Leu Gly Pro Trp Val Pro Leu Ala Ala LeuAla Arg Glu 460 465 470 ttt gat cga ttt ggc acc ata cgc acc ata gac taccga aaa ggt gat 1553 Phe Asp Arg Phe Gly Thr Ile Arg Thr Ile Asp Tyr ArgLys Gly Asp 475 480 485 490 agt tgg gca tat atc cag tat gaa agc ctg gatgca gcg cat gct gcc 1601 Ser Trp Ala Tyr Ile Gln Tyr Glu Ser Leu Asp AlaAla His Ala Ala 495 500 505 tgg acc cat atg cgg ggc ttc cca ctt ggt ggccca gat cga cgc ctt 1649 Trp Thr His Met Arg Gly Phe Pro Leu Gly Gly ProAsp Arg Arg Leu 510 515 520 aga gta gac ttt gcc gac acc gaa cat cgt taccag cag cag tat ctg 1697 Arg Val Asp Phe Ala Asp Thr Glu His Arg Tyr GlnGln Gln Tyr Leu 525 530 535 cag cct ctg ccc ttg act cat tat gag ctg gtgaca gat gct ttt gga 1745 Gln Pro Leu Pro Leu Thr His Tyr Glu Leu Val ThrAsp Ala Phe Gly 540 545 550 cat cgg gca cca gac cct ttg agg ggt gct cgggat agg aca cca ccc 1793 His Arg Ala Pro Asp Pro Leu Arg Gly Ala Arg AspArg Thr Pro Pro 555 560 565 570 tta cta tac aga gat cgt gat agg gac ctttat cct gac tct gat tgg 1841 Leu Leu Tyr Arg Asp Arg Asp Arg Asp Leu TyrPro Asp Ser Asp Trp 575 580 585 gtg cca ccc cca ccc cca gtc cga gaa cgcagc act cgg act gca gct 1889 Val Pro Pro Pro Pro Pro Val Arg Glu Arg SerThr Arg Thr Ala Ala 590 595 600 act tct gtg cct gct tat gag cca ctg gatagc cta gat cgc agg cgg 1937 Thr Ser Val Pro Ala Tyr Glu Pro Leu Asp SerLeu Asp Arg Arg Arg 605 610 615 gat ggt tgg tcc ttg gac cgg gac aga ggtgat cga gat ctg ccc agc 1985 Asp Gly Trp Ser Leu Asp Arg Asp Arg Gly AspArg Asp Leu Pro Ser 620 625 630 agc aga gac cag cct agg aag cga agg ctgcct gag gag agt gga gga 2033 Ser Arg Asp Gln Pro Arg Lys Arg Arg Leu ProGlu Glu Ser Gly Gly 635 640 645 650 cgt cat ctg gat agg tct cct gag agtgac cgc cca cga aaa cgt cac 2081 Arg His Leu Asp Arg Ser Pro Glu Ser AspArg Pro Arg Lys Arg His 655 660 665 tgc gct cct tct cct gac cgc agt ccagaa ttg agc agt agc cgg gat 2129 Cys Ala Pro Ser Pro Asp Arg Ser Pro GluLeu Ser Ser Ser Arg Asp 670 675 680 cgt tac aac agc gac aat gat cga tcttcc cgt ctt ctc ttg gaa agg 2177 Arg Tyr Asn Ser Asp Asn Asp Arg Ser SerArg Leu Leu Leu Glu Arg 685 690 695 ccc tct cca atc aga gac gga cga ggtagt ttg gag aag agc cag ggt 2225 Pro Ser Pro Ile Arg Asp Gly Arg Gly SerLeu Glu Lys Ser Gln Gly 700 705 710 gac aag cga gac cgt aaa aac tct gcatca gct gaa cga gat agg aag 2273 Asp Lys Arg Asp Arg Lys Asn Ser Ala SerAla Glu Arg Asp Arg Lys 715 720 725 730 cac cgg aca act gct ccc act gaggga aaa agc cct ctg aaa aaa gaa 2321 His Arg Thr Thr Ala Pro Thr Glu GlyLys Ser Pro Leu Lys Lys Glu 735 740 745 gac cgc tct gat ggg agt gca cctagc acc agc act gct tcc tcc aag 2369 Asp Arg Ser Asp Gly Ser Ala Pro SerThr Ser Thr Ala Ser Ser Lys 750 755 760 ctg aag tcc ccg tcc cag aaa caggat ggg ggg aca gcc cct gtg gca 2417 Leu Lys Ser Pro Ser Gln Lys Gln AspGly Gly Thr Ala Pro Val Ala 765 770 775 tca gcc tct ccc aaa ctc tgt ttggcc tgg cag ggc atg ctt cta ctg 2465 Ser Ala Ser Pro Lys Leu Cys Leu AlaTrp Gln Gly Met Leu Leu Leu 780 785 790 aag aac agc aac ttt cct tcc aacatg cat ctg ttg cag ggt gac ctc 2513 Lys Asn Ser Asn Phe Pro Ser Asn MetHis Leu Leu Gln Gly Asp Leu 795 800 805 810 caa gtg gct agt agt ctt cttgtg gag ggt tca act gga ggc aaa gtg 2561 Gln Val Ala Ser Ser Leu Leu ValGlu Gly Ser Thr Gly Gly Lys Val 815 820 825 gcc cag ctc aag atc act cagcgt ctc cgt ttg gac cag ccc aag ttg 2609 Ala Gln Leu Lys Ile Thr Gln ArgLeu Arg Leu Asp Gln Pro Lys Leu 830 835 840 gat gaa gta act cga cgc atcaaa gta gca ggg ccc aat ggt tat gcc 2657 Asp Glu Val Thr Arg Arg Ile LysVal Ala Gly Pro Asn Gly Tyr Ala 845 850 855 att ctt ttg gct gtg cct ggaagt tct gac agc cgg tcc tcc tct tcc 2705 Ile Leu Leu Ala Val Pro Gly SerSer Asp Ser Arg Ser Ser Ser Ser 860 865 870 tca gct gca tca gac act gccact tct act cag agg cca ctt agg aac 2753 Ser Ala Ala Ser Asp Thr Ala ThrSer Thr Gln Arg Pro Leu Arg Asn 875 880 885 890 ctt gtg tcc tat tta aagcaa aag cag gca gcc ggg gtg atc agc ctc 2801 Leu Val Ser Tyr Leu Lys GlnLys Gln Ala Ala Gly Val Ile Ser Leu 895 900 905 cct gtg ggg ggc aac aaagac aag gaa aac acc ggg gtc ctt cat gcc 2849 Pro Val Gly Gly Asn Lys AspLys Glu Asn Thr Gly Val Leu His Ala 910 915 920 ttc cca cct tgt gag ttctcc cag cag ttc ctg gat tcc cct gcc aag 2897 Phe Pro Pro Cys Glu Phe SerGln Gln Phe Leu Asp Ser Pro Ala Lys 925 930 935 gca ctg gcc aaa tct gaagaa gat tac ctg gtc atg atc att gtc cgt 2945 Ala Leu Ala Lys Ser Glu GluAsp Tyr Leu Val Met Ile Ile Val Arg 940 945 950 gct ttg aaa agt cca gccgca ttt cat gag cag aga agg agc ttg gag 2993 Ala Leu Lys Ser Pro Ala AlaPhe His Glu Gln Arg Arg Ser Leu Glu 955 960 965 970 cgg gcc agg aca gaggac tat ctc aaa cgg aag att cgt tcc cgg ccg 3041 Arg Ala Arg Thr Glu AspTyr Leu Lys Arg Lys Ile Arg Ser Arg Pro 975 980 985 gag aga tcg gag ctggtc agg atg cac att ttg gaa gag acc tcg gct 3089 Glu Arg Ser Glu Leu ValArg Met His Ile Leu Glu Glu Thr Ser Ala 990 995 1000 gag cca tcc ctc caggcc aag cag ctg aag ctg aag aga gcc aga 3134 Glu Pro Ser Leu Gln Ala LysGln Leu Lys Leu Lys Arg Ala Arg 1005 1010 1015 cta gcc gat gac ctc aatgag aag att gca cag agg cct ggc ccc 3179 Leu Ala Asp Asp Leu Asn Glu LysIle Ala Gln Arg Pro Gly Pro 1020 1025 1030 atg gag ctg gtg gag aag aacatc ctt cct gtt gag tcc agc ctg 3224 Met Glu Leu Val Glu Lys Asn Ile LeuPro Val Glu Ser Ser Leu 1035 1040 1045 aag gaa gcc atc att gtg ggc caggtg aac tat ccc aaa gta gca 3269 Lys Glu Ala Ile Ile Val Gly Gln Val AsnTyr Pro Lys Val Ala 1050 1055 1060 gac agc tct tcc ttc gat gag gac agcagc gat gcc tta tcc ccc 3314 Asp Ser Ser Ser Phe Asp Glu Asp Ser Ser AspAla Leu Ser Pro 1065 1070 1075 gag cag cct gcc agc cat gag tcc cag ggttct gtg ccg tca ccc 3359 Glu Gln Pro Ala Ser His Glu Ser Gln Gly Ser ValPro Ser Pro 1080 1085 1090 ctg gag gcc cga gtc agc gaa cca ctg ctc agtgcc acc tct gca 3404 Leu Glu Ala Arg Val Ser Glu Pro Leu Leu Ser Ala ThrSer Ala 1095 1100 1105 tcc ccc acc cag gtt gtg tct caa ctt ccg atg ggccgg gat tcc 3449 Ser Pro Thr Gln Val Val Ser Gln Leu Pro Met Gly Arg AspSer 1110 1115 1120 aga gaa atg ctt ttc ctg gca gag cag cct cct ctg cctccc cca 3494 Arg Glu Met Leu Phe Leu Ala Glu Gln Pro Pro Leu Pro Pro Pro1125 1130 1135 cct ctg ctg cct ccc agc ctc acc aat gga acc act atc cccact 3539 Pro Leu Leu Pro Pro Ser Leu Thr Asn Gly Thr Thr Ile Pro Thr1140 1145 1150 gcc aag tcc acc ccc aca ctc att aag caa agc caa ccc aagtct 3584 Ala Lys Ser Thr Pro Thr Leu Ile Lys Gln Ser Gln Pro Lys Ser1155 1160 1165 gcc agt gag aag tca cag cgc agc aag aag gcc aag gag ctgaag 3629 Ala Ser Glu Lys Ser Gln Arg Ser Lys Lys Ala Lys Glu Leu Lys1170 1175 1180 cca aag gtg aag aag ctc aag tac cac cag tac atc ccc ccggac 3674 Pro Lys Val Lys Lys Leu Lys Tyr His Gln Tyr Ile Pro Pro Asp1185 1190 1195 cag aag cag gac agg ggg gca ccc ccc atg gac tca tcc tacgcc 3719 Gln Lys Gln Asp Arg Gly Ala Pro Pro Met Asp Ser Ser Tyr Ala1200 1205 1210 aag atc ctg cag cag cag cag ctc ttc ctc cag ctg cag atcctc 3764 Lys Ile Leu Gln Gln Gln Gln Leu Phe Leu Gln Leu Gln Ile Leu1215 1220 1225 aac cag cag cag cag cag cac cac aac tac cag gcc atc ctgcct 3809 Asn Gln Gln Gln Gln Gln His His Asn Tyr Gln Ala Ile Leu Pro1230 1235 1240 gcc ccg cca aag tca gca ggc gag gcc ctg gga agc agc gggacc 3854 Ala Pro Pro Lys Ser Ala Gly Glu Ala Leu Gly Ser Ser Gly Thr1245 1250 1255 ccc cca gta cgc agc ctc tcc act acc aat agc agc tcc agctcg 3899 Pro Pro Val Arg Ser Leu Ser Thr Thr Asn Ser Ser Ser Ser Ser1260 1265 1270 ggc gcc cct ggg ccc tgt ggg ctg gca cgt cag aac agc acctca 3944 Gly Ala Pro Gly Pro Cys Gly Leu Ala Arg Gln Asn Ser Thr Ser1275 1280 1285 ctg act ggc aag ccg gga gcc ctg ccg gcc aac ctg gac gacatg 3989 Leu Thr Gly Lys Pro Gly Ala Leu Pro Ala Asn Leu Asp Asp Met1290 1295 1300 aag gtg gca gag ctg aag cag gag ctg aag ttg cga tca ctgcct 4034 Lys Val Ala Glu Leu Lys Gln Glu Leu Lys Leu Arg Ser Leu Pro1305 1310 1315 gtc tcg ggc acc aaa act gag ctg att gag cgc ctt cga gcctat 4079 Val Ser Gly Thr Lys Thr Glu Leu Ile Glu Arg Leu Arg Ala Tyr1320 1325 1330 caa gac caa atc agc cct gtg cca gga gcc ccc aag gcc cctgcc 4124 Gln Asp Gln Ile Ser Pro Val Pro Gly Ala Pro Lys Ala Pro Ala1335 1340 1345 gcc acc tct atc ctg cac aag gct ggc gag gtg gtg gta gccttc 4169 Ala Thr Ser Ile Leu His Lys Ala Gly Glu Val Val Val Ala Phe1350 1355 1360 cca gcg gcc cgg ctg agc acg ggg cca gcc ctg gtg gca gcaggc 4214 Pro Ala Ala Arg Leu Ser Thr Gly Pro Ala Leu Val Ala Ala Gly1365 1370 1375 ctg gct cca gct gag gtg gtg gtg gcc acg gtg gcc agc agtggg 4259 Leu Ala Pro Ala Glu Val Val Val Ala Thr Val Ala Ser Ser Gly1380 1385 1390 gtg gtg aag ttt ggc agc acg ggc tcc acg ccc ccc gtg tctccc 4304 Val Val Lys Phe Gly Ser Thr Gly Ser Thr Pro Pro Val Ser Pro1395 1400 1405 acc ccc tcg gag cgc tca ctg ctc agc acg ggc gat gaa aactcc 4349 Thr Pro Ser Glu Arg Ser Leu Leu Ser Thr Gly Asp Glu Asn Ser1410 1415 1420 acc ccc ggg gac acc ttt ggt gag atg gtg aca tca cct ctgacg 4394 Thr Pro Gly Asp Thr Phe Gly Glu Met Val Thr Ser Pro Leu Thr1425 1430 1435 cag ctg acc ctg cag gcc tcg cca ctg cag atc ctc gtg aaggag 4439 Gln Leu Thr Leu Gln Ala Ser Pro Leu Gln Ile Leu Val Lys Glu1440 1445 1450 gag ggc ccc cgg gcc ggg tcc tgt tgc ctg agc cct ggg gggcgg 4484 Glu Gly Pro Arg Ala Gly Ser Cys Cys Leu Ser Pro Gly Gly Arg1455 1460 1465 gcg gag cta gag ggg cgc gac aag gac cag atg ctg cag gagaaa 4529 Ala Glu Leu Glu Gly Arg Asp Lys Asp Gln Met Leu Gln Glu Lys1470 1475 1480 gac aag cag atc gag gcg ctg acg cgc atg ctc cgg cag aagcag 4574 Asp Lys Gln Ile Glu Ala Leu Thr Arg Met Leu Arg Gln Lys Gln1485 1490 1495 cag ctg gtg gag cgg ctc aag ctg cag ctg gag cag gag aagcga 4619 Gln Leu Val Glu Arg Leu Lys Leu Gln Leu Glu Gln Glu Lys Arg1500 1505 1510 gcc cag cag ccc gcc ccc gcc ccc gcc ccc ctc ggc acc cccgtg 4664 Ala Gln Gln Pro Ala Pro Ala Pro Ala Pro Leu Gly Thr Pro Val1515 1520 1525 aag cag gag aac agc ttc tcc agc tgc cag ctg agc cag cagccc 4709 Lys Gln Glu Asn Ser Phe Ser Ser Cys Gln Leu Ser Gln Gln Pro1530 1535 1540 ctg ggc ccc gct cac cca ttc aac ccc agc ctg gcg gcc ccagcc 4754 Leu Gly Pro Ala His Pro Phe Asn Pro Ser Leu Ala Ala Pro Ala1545 1550 1555 acc aac cac ata gac cct tgt gct gtg gcc ccg ggg ccc ccgtcc 4799 Thr Asn His Ile Asp Pro Cys Ala Val Ala Pro Gly Pro Pro Ser1560 1565 1570 gtg gtg gtg aag cag gaa gcc ttg cag cct gag ccc gag ccggtc 4844 Val Val Val Lys Gln Glu Ala Leu Gln Pro Glu Pro Glu Pro Val1575 1580 1585 ccc gcc ccc cag ttg ctt ctg ggg cct cag ggc ccc agc ctcatc 4889 Pro Ala Pro Gln Leu Leu Leu Gly Pro Gln Gly Pro Ser Leu Ile1590 1595 1600 aag ggg gtt gca cct ccc acc ctc atc acc gac tcc aca gggacc 4934 Lys Gly Val Ala Pro Pro Thr Leu Ile Thr Asp Ser Thr Gly Thr1605 1610 1615 cac ctt gtc ctc acc gtg acc aat aag aat gca gac agc cctggc 4979 His Leu Val Leu Thr Val Thr Asn Lys Asn Ala Asp Ser Pro Gly1620 1625 1630 ctg tcc agt ggg agc ccc cag cag ccc tcg tcc cag cct ggctct 5024 Leu Ser Ser Gly Ser Pro Gln Gln Pro Ser Ser Gln Pro Gly Ser1635 1640 1645 cca gcg cct gcc ccc tct gcc cag atg gac ctg gag cac ccactg 5069 Pro Ala Pro Ala Pro Ser Ala Gln Met Asp Leu Glu His Pro Leu1650 1655 1660 cag ccc ctc ttt ggg acc ccc act tct ctg ctg aag aag gaacca 5114 Gln Pro Leu Phe Gly Thr Pro Thr Ser Leu Leu Lys Lys Glu Pro1665 1670 1675 cct ggc tat gag gaa gcc atg agc cag cag ccc aaa cag caggaa 5159 Pro Gly Tyr Glu Glu Ala Met Ser Gln Gln Pro Lys Gln Gln Glu1680 1685 1690 aat ggt tcc tca agc cag cag atg gac gac ctg ttt gac attctc 5204 Asn Gly Ser Ser Ser Gln Gln Met Asp Asp Leu Phe Asp Ile Leu1695 1700 1705 att cag agc gga gaa att tca gca gat ttc aag gag ccg ccatcc 5249 Ile Gln Ser Gly Glu Ile Ser Ala Asp Phe Lys Glu Pro Pro Ser1710 1715 1720 ctg cca ggg aag gag aag cca tcc ccg aag aca gtc tgt gggtcc 5294 Leu Pro Gly Lys Glu Lys Pro Ser Pro Lys Thr Val Cys Gly Ser1725 1730 1735 ccc ctg gca gca cag cca tca cct tct gct gag ctc ccc caggct 5339 Pro Leu Ala Ala Gln Pro Ser Pro Ser Ala Glu Leu Pro Gln Ala1740 1745 1750 gcc cca cct cct cca ggc tca ccc tcc ctc cct gga cgc ctggag 5384 Ala Pro Pro Pro Pro Gly Ser Pro Ser Leu Pro Gly Arg Leu Glu1755 1760 1765 gac ttc ctg gag agc agc acg ggg ctg ccc ctg ctg acc agtggg 5429 Asp Phe Leu Glu Ser Ser Thr Gly Leu Pro Leu Leu Thr Ser Gly1770 1775 1780 cat gac ggg cca gag ccc ctt tcc ctc att gac gac ctc catagc 5474 His Asp Gly Pro Glu Pro Leu Ser Leu Ile Asp Asp Leu His Ser1785 1790 1795 cag atg ctg agc agc act gcc atc ctg gac cac ccc ccg tcaccc 5519 Gln Met Leu Ser Ser Thr Ala Ile Leu Asp His Pro Pro Ser Pro1800 1805 1810 atg gac acc tcg gaa ttg cac ttt gtt cct gag ccc agc agcacc 5564 Met Asp Thr Ser Glu Leu His Phe Val Pro Glu Pro Ser Ser Thr1815 1820 1825 atg ggc ctg gac ctg gct gat ggc cac ctg gac agc atg gactgg 5609 Met Gly Leu Asp Leu Ala Asp Gly His Leu Asp Ser Met Asp Trp1830 1835 1840 ctg gag ctg tcg tca ggt ggt ccc gtg ctg agc cta gcc cccctc 5654 Leu Glu Leu Ser Ser Gly Gly Pro Val Leu Ser Leu Ala Pro Leu1845 1850 1855 agc acc aca gcc ccc agc ctc ttc tcc aca gac ttc ctc gatggc 5699 Ser Thr Thr Ala Pro Ser Leu Phe Ser Thr Asp Phe Leu Asp Gly1860 1865 1870 cat gat ttg cag ctg cac tgg gat tcc tgc ttg tagctctctg5742 His Asp Leu Gln Leu His Trp Asp Ser Cys Leu 1875 1880 gctcaagacggggtggggaa ggggctggga gccagggtac tccaatgcgt ggctctcctg 5802 cgtgattcggcctctccaca tggttgtgag tcttgacaat cacagcccct gctttttccc 5862 ttccctgggaggctagaaca gagaagccct tactcctggt tcagtgccac gcagggcaga 5922 ggagagcagctgtcaagaag cagccctggc tctcacgctg gggttttgga cacacggtca 5982 gggtcagggccatttcagct tgacctcctt ttttgaggtc agggggcact gtctgtctgg 6042 ctacaatttggctaaggtag gtgaagcctg gccaggcggg aggcttctct tctgacccag 6102 ggctgagacaggttaagggg tgaatctcct tcctttctct ccctgctttg ctgtgaaggg 6162 agaaattagcctgggcctct accccctatt ccctgtgtct gccaacccca ggatcccagg 6222 gctccctgccattttagtgt cttggtgtag tgtaaccatt tagtggttgg tggcaacaat 6282 tttatgtacaggtgtatata cctctatatt atatatcgac atacatatat atttttgggg 6342 gggggcggacaggagatggg tgcaactccc tcccatccta ctctcacaga agggcctgga 6402 tgcaaggttacccttgagct gtgtgccaca gtctggtgcc cagtctggca tgcagctacc 6462 caggcccacccatcacgtgt gattgacatg taggtaccct gccacggcct atgccccacc 6522 tgccctgcttcctggctcct tatcagtgcc atgagggcag aggtgctacc tggccttcct 6582 gccaggagctctccacccac tcacattccg tccccgccgc ctcactgcag ccagcgtggt 6642 cctaggacaggaggagcttc gggcccagct tcaccctgcg gtggggctga ggggtggcca 6702 tctcctgccctggggccact ggcttcacat tctgggctga ctcatagggg agtaggggtg 6762 gagtcaccaaaaccagtgct gggacaaaga tggggaaggt gtgtgaactt tttaaaataa 6822 acacaaaaacacag 6836 2 1883 PRT Homo sapiens 2 Met Arg Thr Ala Gly Arg Asp Pro ValPro Arg Arg Ser Pro Arg Trp 1 5 10 15 Arg Arg Ala Val Pro Leu Cys GluThr Ser Ala Gly Arg Arg Val Thr 20 25 30 Gln Leu Arg Gly Asp Asp Leu ArgArg Pro Ala Thr Met Lys Gly Lys 35 40 45 Glu Arg Ser Pro Val Lys Ala LysArg Ser Arg Gly Gly Glu Asp Ser 50 55 60 Thr Ser Arg Gly Glu Arg Ser LysLys Leu Gly Gly Ser Gly Gly Ser 65 70 75 80 Asn Gly Ser Ser Ser Gly LysThr Asp Ser Gly Gly Gly Ser Arg Arg 85 90 95 Ser Leu Leu Leu Asp Lys SerSer Ser Arg Gly Gly Ser Arg Glu Tyr 100 105 110 Asp Thr Gly Gly Gly SerSer Ser Ser Arg Leu His Ser Tyr Ser Ser 115 120 125 Pro Ser Thr Lys AsnSer Ser Gly Gly Gly Glu Ser Arg Ser Ser Ser 130 135 140 Arg Gly Gly GlyGly Glu Ser Arg Ser Ser Gly Ala Ala Ser Ser Ala 145 150 155 160 Pro GlyGly Gly Asp Gly Ala Glu Tyr Lys Thr Leu Lys Ile Ser Glu 165 170 175 LeuGly Ser Gln Leu Ser Asp Glu Ala Val Glu Asp Gly Leu Phe His 180 185 190Glu Phe Lys Arg Phe Gly Asp Val Ser Val Lys Ile Ser His Leu Ser 195 200205 Gly Ser Gly Ser Gly Asp Glu Arg Val Ala Phe Val Asn Phe Arg Arg 210215 220 Pro Glu Asp Ala Arg Ala Ala Lys His Ala Arg Gly Arg Leu Val Leu225 230 235 240 Tyr Asp Arg Pro Leu Lys Ile Glu Ala Val Tyr Val Ser ArgArg Arg 245 250 255 Ser Arg Ser Pro Leu Asp Lys Asp Thr Tyr Pro Pro SerAla Ser Val 260 265 270 Val Gly Ala Ser Val Gly Gly His Arg His Pro ProGly Gly Gly Gly 275 280 285 Gly Gln Arg Ser Leu Ser Pro Gly Gly Ala AlaLeu Gly Tyr Arg Asp 290 295 300 Tyr Arg Leu Gln Gln Leu Ala Leu Gly ArgLeu Pro Pro Pro Pro Pro 305 310 315 320 Pro Pro Leu Pro Arg Asp Leu GluArg Glu Arg Asp Tyr Pro Phe Tyr 325 330 335 Glu Arg Val Arg Pro Ala TyrSer Leu Glu Pro Arg Val Gly Ala Gly 340 345 350 Ala Gly Ala Ala Pro PheArg Glu Val Asp Glu Ile Ser Pro Glu Asp 355 360 365 Asp Gln Arg Ala AsnArg Thr Leu Phe Leu Gly Asn Leu Asp Ile Thr 370 375 380 Val Thr Glu SerAsp Leu Arg Arg Ala Phe Asp Arg Phe Gly Val Ile 385 390 395 400 Thr GluVal Asp Ile Lys Arg Pro Ser Arg Gly Gln Thr Ser Thr Tyr 405 410 415 GlyPhe Leu Lys Phe Glu Asn Leu Asp Met Ser His Arg Ala Lys Leu 420 425 430Ala Met Ser Gly Lys Ile Ile Ile Arg Asn Pro Ile Lys Ile Gly Tyr 435 440445 Gly Lys Ala Thr Pro Thr Thr Arg Leu Trp Val Gly Gly Leu Gly Pro 450455 460 Trp Val Pro Leu Ala Ala Leu Ala Arg Glu Phe Asp Arg Phe Gly Thr465 470 475 480 Ile Arg Thr Ile Asp Tyr Arg Lys Gly Asp Ser Trp Ala TyrIle Gln 485 490 495 Tyr Glu Ser Leu Asp Ala Ala His Ala Ala Trp Thr HisMet Arg Gly 500 505 510 Phe Pro Leu Gly Gly Pro Asp Arg Arg Leu Arg ValAsp Phe Ala Asp 515 520 525 Thr Glu His Arg Tyr Gln Gln Gln Tyr Leu GlnPro Leu Pro Leu Thr 530 535 540 His Tyr Glu Leu Val Thr Asp Ala Phe GlyHis Arg Ala Pro Asp Pro 545 550 555 560 Leu Arg Gly Ala Arg Asp Arg ThrPro Pro Leu Leu Tyr Arg Asp Arg 565 570 575 Asp Arg Asp Leu Tyr Pro AspSer Asp Trp Val Pro Pro Pro Pro Pro 580 585 590 Val Arg Glu Arg Ser ThrArg Thr Ala Ala Thr Ser Val Pro Ala Tyr 595 600 605 Glu Pro Leu Asp SerLeu Asp Arg Arg Arg Asp Gly Trp Ser Leu Asp 610 615 620 Arg Asp Arg GlyAsp Arg Asp Leu Pro Ser Ser Arg Asp Gln Pro Arg 625 630 635 640 Lys ArgArg Leu Pro Glu Glu Ser Gly Gly Arg His Leu Asp Arg Ser 645 650 655 ProGlu Ser Asp Arg Pro Arg Lys Arg His Cys Ala Pro Ser Pro Asp 660 665 670Arg Ser Pro Glu Leu Ser Ser Ser Arg Asp Arg Tyr Asn Ser Asp Asn 675 680685 Asp Arg Ser Ser Arg Leu Leu Leu Glu Arg Pro Ser Pro Ile Arg Asp 690695 700 Gly Arg Gly Ser Leu Glu Lys Ser Gln Gly Asp Lys Arg Asp Arg Lys705 710 715 720 Asn Ser Ala Ser Ala Glu Arg Asp Arg Lys His Arg Thr ThrAla Pro 725 730 735 Thr Glu Gly Lys Ser Pro Leu Lys Lys Glu Asp Arg SerAsp Gly Ser 740 745 750 Ala Pro Ser Thr Ser Thr Ala Ser Ser Lys Leu LysSer Pro Ser Gln 755 760 765 Lys Gln Asp Gly Gly Thr Ala Pro Val Ala SerAla Ser Pro Lys Leu 770 775 780 Cys Leu Ala Trp Gln Gly Met Leu Leu LeuLys Asn Ser Asn Phe Pro 785 790 795 800 Ser Asn Met His Leu Leu Gln GlyAsp Leu Gln Val Ala Ser Ser Leu 805 810 815 Leu Val Glu Gly Ser Thr GlyGly Lys Val Ala Gln Leu Lys Ile Thr 820 825 830 Gln Arg Leu Arg Leu AspGln Pro Lys Leu Asp Glu Val Thr Arg Arg 835 840 845 Ile Lys Val Ala GlyPro Asn Gly Tyr Ala Ile Leu Leu Ala Val Pro 850 855 860 Gly Ser Ser AspSer Arg Ser Ser Ser Ser Ser Ala Ala Ser Asp Thr 865 870 875 880 Ala ThrSer Thr Gln Arg Pro Leu Arg Asn Leu Val Ser Tyr Leu Lys 885 890 895 GlnLys Gln Ala Ala Gly Val Ile Ser Leu Pro Val Gly Gly Asn Lys 900 905 910Asp Lys Glu Asn Thr Gly Val Leu His Ala Phe Pro Pro Cys Glu Phe 915 920925 Ser Gln Gln Phe Leu Asp Ser Pro Ala Lys Ala Leu Ala Lys Ser Glu 930935 940 Glu Asp Tyr Leu Val Met Ile Ile Val Arg Ala Leu Lys Ser Pro Ala945 950 955 960 Ala Phe His Glu Gln Arg Arg Ser Leu Glu Arg Ala Arg ThrGlu Asp 965 970 975 Tyr Leu Lys Arg Lys Ile Arg Ser Arg Pro Glu Arg SerGlu Leu Val 980 985 990 Arg Met His Ile Leu Glu Glu Thr Ser Ala Glu ProSer Leu Gln Ala 995 1000 1005 Lys Gln Leu Lys Leu Lys Arg Ala Arg LeuAla Asp Asp Leu Asn 1010 1015 1020 Glu Lys Ile Ala Gln Arg Pro Gly ProMet Glu Leu Val Glu Lys 1025 1030 1035 Asn Ile Leu Pro Val Glu Ser SerLeu Lys Glu Ala Ile Ile Val 1040 1045 1050 Gly Gln Val Asn Tyr Pro LysVal Ala Asp Ser Ser Ser Phe Asp 1055 1060 1065 Glu Asp Ser Ser Asp AlaLeu Ser Pro Glu Gln Pro Ala Ser His 1070 1075 1080 Glu Ser Gln Gly SerVal Pro Ser Pro Leu Glu Ala Arg Val Ser 1085 1090 1095 Glu Pro Leu LeuSer Ala Thr Ser Ala Ser Pro Thr Gln Val Val 1100 1105 1110 Ser Gln LeuPro Met Gly Arg Asp Ser Arg Glu Met Leu Phe Leu 1115 1120 1125 Ala GluGln Pro Pro Leu Pro Pro Pro Pro Leu Leu Pro Pro Ser 1130 1135 1140 LeuThr Asn Gly Thr Thr Ile Pro Thr Ala Lys Ser Thr Pro Thr 1145 1150 1155Leu Ile Lys Gln Ser Gln Pro Lys Ser Ala Ser Glu Lys Ser Gln 1160 11651170 Arg Ser Lys Lys Ala Lys Glu Leu Lys Pro Lys Val Lys Lys Leu 11751180 1185 Lys Tyr His Gln Tyr Ile Pro Pro Asp Gln Lys Gln Asp Arg Gly1190 1195 1200 Ala Pro Pro Met Asp Ser Ser Tyr Ala Lys Ile Leu Gln GlnGln 1205 1210 1215 Gln Leu Phe Leu Gln Leu Gln Ile Leu Asn Gln Gln GlnGln Gln 1220 1225 1230 His His Asn Tyr Gln Ala Ile Leu Pro Ala Pro ProLys Ser Ala 1235 1240 1245 Gly Glu Ala Leu Gly Ser Ser Gly Thr Pro ProVal Arg Ser Leu 1250 1255 1260 Ser Thr Thr Asn Ser Ser Ser Ser Ser GlyAla Pro Gly Pro Cys 1265 1270 1275 Gly Leu Ala Arg Gln Asn Ser Thr SerLeu Thr Gly Lys Pro Gly 1280 1285 1290 Ala Leu Pro Ala Asn Leu Asp AspMet Lys Val Ala Glu Leu Lys 1295 1300 1305 Gln Glu Leu Lys Leu Arg SerLeu Pro Val Ser Gly Thr Lys Thr 1310 1315 1320 Glu Leu Ile Glu Arg LeuArg Ala Tyr Gln Asp Gln Ile Ser Pro 1325 1330 1335 Val Pro Gly Ala ProLys Ala Pro Ala Ala Thr Ser Ile Leu His 1340 1345 1350 Lys Ala Gly GluVal Val Val Ala Phe Pro Ala Ala Arg Leu Ser 1355 1360 1365 Thr Gly ProAla Leu Val Ala Ala Gly Leu Ala Pro Ala Glu Val 1370 1375 1380 Val ValAla Thr Val Ala Ser Ser Gly Val Val Lys Phe Gly Ser 1385 1390 1395 ThrGly Ser Thr Pro Pro Val Ser Pro Thr Pro Ser Glu Arg Ser 1400 1405 1410Leu Leu Ser Thr Gly Asp Glu Asn Ser Thr Pro Gly Asp Thr Phe 1415 14201425 Gly Glu Met Val Thr Ser Pro Leu Thr Gln Leu Thr Leu Gln Ala 14301435 1440 Ser Pro Leu Gln Ile Leu Val Lys Glu Glu Gly Pro Arg Ala Gly1445 1450 1455 Ser Cys Cys Leu Ser Pro Gly Gly Arg Ala Glu Leu Glu GlyArg 1460 1465 1470 Asp Lys Asp Gln Met Leu Gln Glu Lys Asp Lys Gln IleGlu Ala 1475 1480 1485 Leu Thr Arg Met Leu Arg Gln Lys Gln Gln Leu ValGlu Arg Leu 1490 1495 1500 Lys Leu Gln Leu Glu Gln Glu Lys Arg Ala GlnGln Pro Ala Pro 1505 1510 1515 Ala Pro Ala Pro Leu Gly Thr Pro Val LysGln Glu Asn Ser Phe 1520 1525 1530 Ser Ser Cys Gln Leu Ser Gln Gln ProLeu Gly Pro Ala His Pro 1535 1540 1545 Phe Asn Pro Ser Leu Ala Ala ProAla Thr Asn His Ile Asp Pro 1550 1555 1560 Cys Ala Val Ala Pro Gly ProPro Ser Val Val Val Lys Gln Glu 1565 1570 1575 Ala Leu Gln Pro Glu ProGlu Pro Val Pro Ala Pro Gln Leu Leu 1580 1585 1590 Leu Gly Pro Gln GlyPro Ser Leu Ile Lys Gly Val Ala Pro Pro 1595 1600 1605 Thr Leu Ile ThrAsp Ser Thr Gly Thr His Leu Val Leu Thr Val 1610 1615 1620 Thr Asn LysAsn Ala Asp Ser Pro Gly Leu Ser Ser Gly Ser Pro 1625 1630 1635 Gln GlnPro Ser Ser Gln Pro Gly Ser Pro Ala Pro Ala Pro Ser 1640 1645 1650 AlaGln Met Asp Leu Glu His Pro Leu Gln Pro Leu Phe Gly Thr 1655 1660 1665Pro Thr Ser Leu Leu Lys Lys Glu Pro Pro Gly Tyr Glu Glu Ala 1670 16751680 Met Ser Gln Gln Pro Lys Gln Gln Glu Asn Gly Ser Ser Ser Gln 16851690 1695 Gln Met Asp Asp Leu Phe Asp Ile Leu Ile Gln Ser Gly Glu Ile1700 1705 1710 Ser Ala Asp Phe Lys Glu Pro Pro Ser Leu Pro Gly Lys GluLys 1715 1720 1725 Pro Ser Pro Lys Thr Val Cys Gly Ser Pro Leu Ala AlaGln Pro 1730 1735 1740 Ser Pro Ser Ala Glu Leu Pro Gln Ala Ala Pro ProPro Pro Gly 1745 1750 1755 Ser Pro Ser Leu Pro Gly Arg Leu Glu Asp PheLeu Glu Ser Ser 1760 1765 1770 Thr Gly Leu Pro Leu Leu Thr Ser Gly HisAsp Gly Pro Glu Pro 1775 1780 1785 Leu Ser Leu Ile Asp Asp Leu His SerGln Met Leu Ser Ser Thr 1790 1795 1800 Ala Ile Leu Asp His Pro Pro SerPro Met Asp Thr Ser Glu Leu 1805 1810 1815 His Phe Val Pro Glu Pro SerSer Thr Met Gly Leu Asp Leu Ala 1820 1825 1830 Asp Gly His Leu Asp SerMet Asp Trp Leu Glu Leu Ser Ser Gly 1835 1840 1845 Gly Pro Val Leu SerLeu Ala Pro Leu Ser Thr Thr Ala Pro Ser 1850 1855 1860 Leu Phe Ser ThrAsp Phe Leu Asp Gly His Asp Leu Gln Leu His 1865 1870 1875 Trp Asp SerCys Leu 1880 3 923 DNA Homo sapiens CDS (551)..(601) 3 tcccatctttcccaccctct tggttgccgc tggccacacg ccctccgctg gcggcgactt 60 ctcagctccgtgcgcccggg ctggacagtg agcctcgaga ggagacgcgg gcggctagag 120 ccggagtggggcgagccgcg gaacccggcc gggagccgcg cgaggcgtga tcggagggta 180 tggttggcatggaattgaat ttcatctgtc tgtgggaatt gtaagcaaga ttgccatcac 240 gaaagccaaagtggatttct ccagtgtggt gtgcctgccc ccttccgtca ttgctgtgaa 300 tgggctggacggaggagggg ccggcgaaaa tgatgatgaa ccagtgctcg tgtccttatc 360 tgcggcacccagtccccaga gtgaagctgt tgccaatgaa ctgcaggagc tctccttgca 420 gcccaagctgaccctaggcc tccaccctgg caggaatccc aatttgcctc cacttagtga 480 gcggaagaatgtgctacagt tgaaactcca gcagcgccgg acccgggaag aactggtgag 540 ccaagggatcatg ccg cca aaa ctg gtt gaa cag cgg atg aag ata tgg 589 Met Pro Pro LysLeu Val Glu Gln Arg Met Lys Ile Trp 1 5 10 aat tca aag ctc taatggacctttttgaagag aagttgtggc ttatgtggag 641 Asn Ser Lys Leu 15 tttacatgggcctctgatgg aagaaagcta atctgtttag tatttgtgca ttttactaaa 701 atggcagcttaaagttgtgt atctgctatt gtgatgccaa tgccggtgtt ttaagtggaa 761 aaaaaatgacctctttgatt tgtgctgtgt acacaagatt tctggaaaag taaagaaaaa 821 ccctttttatggctcacaca gcttaagagt agctgtctct caaacgtgcg ctcacagttg 881 agctgcttttgttttattct aaataaattg tttcttttga gg 923 4 17 PRT Homo sapiens 4 Met ProPro Lys Leu Val Glu Gln Arg Met Lys Ile Trp Asn Ser Lys 1 5 10 15 Leu 51034 DNA Homo sapiens CDS (551)..(625) 5 tcccatcttt cccaccctcttggttgccgc tggccacacg ccctccgctg gcggcgactt 60 ctcagctccg tgcgcccgggctggacagtg agcctcgaga ggagacgcgg gcggctagag 120 ccggagtggg gcgagccgcggaacccggcc gggagccgcg cgaggcgtga tcggagggta 180 tggttggcat ggaattgaatttcatctgtc tgtgggaatt gtaagcaaga ttgccatcac 240 gaaagccaaa gtggatttctccagtgtggt gtgcctgccc ccttccgtca ttgctgtgaa 300 tgggctggac ggaggaggggccggcgaaaa tgatgatgaa ccagtgctcg tgtccttatc 360 tgcggcaccc agtccccagagtgaagctgt tgccaatgaa ctgcaggagc tctccttgca 420 gcccaagctg accctaggcctccaccctgg caggaatccc aatttgcctc cacttagtga 480 gcggaagaat gtgctacagttgaaactcca gcagcgccgg acccgggaag aactggtgag 540 ccaagggatc atg ccg cggttt ggt ttt cag ata gga gtt agg tat gag 589 Met Pro Arg Phe Gly Phe GlnIle Gly Val Arg Tyr Glu 1 5 10 aac aag aag aga gaa aac ttg gcg ctg accctg tta tagtggttat 635 Asn Lys Lys Arg Glu Asn Leu Ala Leu Thr Leu Leu15 20 25 agtggtgtcc ctaaagggag gaaatgattt cagcaaaact ggttgaacagcggatgaaga 695 tatggaattc aaagctctaa tggacctttt tgaagagaag ttgtggcttatgtggagttt 755 acatgggcct ctgatggaag aaagctaatc tgtttagtat ttgtgcattttactaaaatg 815 gcagcttaaa gttgtgtatc tgctattgtg atgccaatgc cggtgttttaagtggaaaaa 875 aaatgacctc tttgatttgt gctgtgtaca caagatttct ggaaaagtaaagaaaaaccc 935 tttttatggc tcacacagct taagagtagc tgtctctcaa acgtgcgctcacagttgagc 995 tgcttttgtt ttattctaaa taaattgttt cttttgagg 1034 6 25 PRTHomo sapiens 6 Met Pro Arg Phe Gly Phe Gln Ile Gly Val Arg Tyr Glu AsnLys Lys 1 5 10 15 Arg Glu Asn Leu Ala Leu Thr Leu Leu 20 25 7 3312 DNAHomo sapiens CDS (84)..(2990) 7 ggcgccttcc tggagcgcgg ggagatgtaaagatagacaa ataattttcc caatgagact 60 gtagaagaga gagcaattgg cca atg aggact gcg ggg cgg gac cct gtg ccg 113 Met Arg Thr Ala Gly Arg Asp Pro ValPro 1 5 10 cgg cgg agt cca aga tgg cgg cgt gcg gtt ccg ctg tgt gaa acgagc 161 Arg Arg Ser Pro Arg Trp Arg Arg Ala Val Pro Leu Cys Glu Thr Ser15 20 25 gcg ggg cgg cgg gtt act cag ctc cgc gga gac gac ctc cga cga ccc209 Ala Gly Arg Arg Val Thr Gln Leu Arg Gly Asp Asp Leu Arg Arg Pro 3035 40 gca aca atg aag gga aaa gag cgc tcg cca gtg aag gcc aaa cgc tcc257 Ala Thr Met Lys Gly Lys Glu Arg Ser Pro Val Lys Ala Lys Arg Ser 4550 55 cgt ggt ggt gag gac tcg act tcc cgc ggt gag cgg agc aag aag tta305 Arg Gly Gly Glu Asp Ser Thr Ser Arg Gly Glu Arg Ser Lys Lys Leu 6065 70 ggg ggc tct ggt ggc agc aat ggg agc agc agc gga aag acc gat agc353 Gly Gly Ser Gly Gly Ser Asn Gly Ser Ser Ser Gly Lys Thr Asp Ser 7580 85 90 ggc ggt ggg tcg cgg cgg agt ctc ctc ctg gac aag tcc agc agt cga401 Gly Gly Gly Ser Arg Arg Ser Leu Leu Leu Asp Lys Ser Ser Ser Arg 95100 105 ggt ggc agc cgc gag tat gat acc ggt ggg ggc agc tcc agt agc cgc449 Gly Gly Ser Arg Glu Tyr Asp Thr Gly Gly Gly Ser Ser Ser Ser Arg 110115 120 ttg cat agt tat agc tcc ccg agc acc aaa aat tct tcg ggc ggg ggc497 Leu His Ser Tyr Ser Ser Pro Ser Thr Lys Asn Ser Ser Gly Gly Gly 125130 135 gag tcg cgc agc agc tcc cgg ggt gga ggc ggg gag tca cgt tcc tct545 Glu Ser Arg Ser Ser Ser Arg Gly Gly Gly Gly Glu Ser Arg Ser Ser 140145 150 ggg gcc gcc tcc tca gct ccc ggc ggc ggg gac ggc gcg gaa tac aag593 Gly Ala Ala Ser Ser Ala Pro Gly Gly Gly Asp Gly Ala Glu Tyr Lys 155160 165 170 act ctg aag ata agc gag ttg ggg tcc cag ctt agt gac gaa gcggtg 641 Thr Leu Lys Ile Ser Glu Leu Gly Ser Gln Leu Ser Asp Glu Ala Val175 180 185 gag gac ggc ctg ttt cat gag ttc aaa cgc ttc ggt gat gta agtgtg 689 Glu Asp Gly Leu Phe His Glu Phe Lys Arg Phe Gly Asp Val Ser Val190 195 200 aaa atc agt cat ctg tcg ggt tct ggc agc ggg gat gag cgg gtagcc 737 Lys Ile Ser His Leu Ser Gly Ser Gly Ser Gly Asp Glu Arg Val Ala205 210 215 ttt gtg aac ttc cgg cgg cca gag gac gcg cgg gcg gcc aag catgcc 785 Phe Val Asn Phe Arg Arg Pro Glu Asp Ala Arg Ala Ala Lys His Ala220 225 230 aga ggc cgc ctg gtg ctc tat gac cgg cct ctg aag ata gaa gctgtg 833 Arg Gly Arg Leu Val Leu Tyr Asp Arg Pro Leu Lys Ile Glu Ala Val235 240 245 250 tat gtg agc cgg cgc cgc agc cgc tcc cct tta gac aaa gatact tat 881 Tyr Val Ser Arg Arg Arg Ser Arg Ser Pro Leu Asp Lys Asp ThrTyr 255 260 265 cct cca tca gcc agt gtg gtc ggg gcc tct gta ggt ggt caccgg cac 929 Pro Pro Ser Ala Ser Val Val Gly Ala Ser Val Gly Gly His ArgHis 270 275 280 ccc cct gga ggt ggt gga ggc cag aga tca ctt tcc cct ggtggc gct 977 Pro Pro Gly Gly Gly Gly Gly Gln Arg Ser Leu Ser Pro Gly GlyAla 285 290 295 gct ttg gga tac aga gac tac cgg ctg cag cag ttg gct cttggc cgc 1025 Ala Leu Gly Tyr Arg Asp Tyr Arg Leu Gln Gln Leu Ala Leu GlyArg 300 305 310 ctg ccc cct cca cct ccg cca cca ttg cct cga gac ctg gagaga gaa 1073 Leu Pro Pro Pro Pro Pro Pro Pro Leu Pro Arg Asp Leu Glu ArgGlu 315 320 325 330 aga gac tac ccg ttc tat gag aga gtg cgc cct gca tacagt ctt gag 1121 Arg Asp Tyr Pro Phe Tyr Glu Arg Val Arg Pro Ala Tyr SerLeu Glu 335 340 345 cca agg gtg gga gct gga gca ggt gct gct cct ttc agagaa gtg gat 1169 Pro Arg Val Gly Ala Gly Ala Gly Ala Ala Pro Phe Arg GluVal Asp 350 355 360 gag att tca ccc gag gat gat cag cga gct aac cgg acgctc ttc ttg 1217 Glu Ile Ser Pro Glu Asp Asp Gln Arg Ala Asn Arg Thr LeuPhe Leu 365 370 375 ggc aac cta gac atc act gta acg gag agt gat tta agaagg gcg ttt 1265 Gly Asn Leu Asp Ile Thr Val Thr Glu Ser Asp Leu Arg ArgAla Phe 380 385 390 gat cgc ttt gga gtc atc aca gaa gta gat atc aag aggcct tct cgc 1313 Asp Arg Phe Gly Val Ile Thr Glu Val Asp Ile Lys Arg ProSer Arg 395 400 405 410 ggc cag act agt act tac ggc ttt ctc aaa ttt gagaac tta gat atg 1361 Gly Gln Thr Ser Thr Tyr Gly Phe Leu Lys Phe Glu AsnLeu Asp Met 415 420 425 tct cac cgg gcc aaa tta gca atg tct ggc aaa attata att cgg aat 1409 Ser His Arg Ala Lys Leu Ala Met Ser Gly Lys Ile IleIle Arg Asn 430 435 440 cct atc aaa att ggt tat ggt aaa gct aca ccc accacc cgc ctc tgg 1457 Pro Ile Lys Ile Gly Tyr Gly Lys Ala Thr Pro Thr ThrArg Leu Trp 445 450 455 gtg gga ggc ctg gga cct tgg gtt cct ctt gct gccctg gca cga gaa 1505 Val Gly Gly Leu Gly Pro Trp Val Pro Leu Ala Ala LeuAla Arg Glu 460 465 470 ttt gat cga ttt ggc acc ata cgc acc ata gac taccga aaa ggt gat 1553 Phe Asp Arg Phe Gly Thr Ile Arg Thr Ile Asp Tyr ArgLys Gly Asp 475 480 485 490 agt tgg gca tat atc cag tat gaa agc ctg gatgca gcg cat gct gcc 1601 Ser Trp Ala Tyr Ile Gln Tyr Glu Ser Leu Asp AlaAla His Ala Ala 495 500 505 tgg acc cat atg cgg ggc ttc cca ctt ggt ggccca gat cga cgc ctt 1649 Trp Thr His Met Arg Gly Phe Pro Leu Gly Gly ProAsp Arg Arg Leu 510 515 520 aga gta gac ttt gcc gac acc gaa cat cgt taccag cag cag tat ctg 1697 Arg Val Asp Phe Ala Asp Thr Glu His Arg Tyr GlnGln Gln Tyr Leu 525 530 535 cag cct ctg ccc ttg act cat tat gag ctg gtgaca gat gct ttt gga 1745 Gln Pro Leu Pro Leu Thr His Tyr Glu Leu Val ThrAsp Ala Phe Gly 540 545 550 cat cgg gca cca gac cct ttg agg ggt gct cgggat agg aca cca ccc 1793 His Arg Ala Pro Asp Pro Leu Arg Gly Ala Arg AspArg Thr Pro Pro 555 560 565 570 tta cta tac aga gat cgt gat agg gac ctttat cct gac tct gat tgg 1841 Leu Leu Tyr Arg Asp Arg Asp Arg Asp Leu TyrPro Asp Ser Asp Trp 575 580 585 gtg cca ccc cca ccc cca gtc cga gaa cgcagc act cgg act gca gct 1889 Val Pro Pro Pro Pro Pro Val Arg Glu Arg SerThr Arg Thr Ala Ala 590 595 600 act tct gtg cct gct tat gag cca ctg gatagc cta gat cgc agg cgg 1937 Thr Ser Val Pro Ala Tyr Glu Pro Leu Asp SerLeu Asp Arg Arg Arg 605 610 615 gat ggt tgg tcc ttg gac cgg gac aga ggtgat cga gat ctg ccc agc 1985 Asp Gly Trp Ser Leu Asp Arg Asp Arg Gly AspArg Asp Leu Pro Ser 620 625 630 agc aga gac cag cct agg aag cga agg ctgcct gag gag agt gga gga 2033 Ser Arg Asp Gln Pro Arg Lys Arg Arg Leu ProGlu Glu Ser Gly Gly 635 640 645 650 cgt cat ctg gat agg tct cct gag agtgac cgc cca cga aaa cgt cac 2081 Arg His Leu Asp Arg Ser Pro Glu Ser AspArg Pro Arg Lys Arg His 655 660 665 tgc gct cct tct cct gac cgc agt ccagaa ttg agc agt agc cgg gat 2129 Cys Ala Pro Ser Pro Asp Arg Ser Pro GluLeu Ser Ser Ser Arg Asp 670 675 680 cgt tac aac agc gac aat gat cga tcttcc cgt ctt ctc ttg gaa agg 2177 Arg Tyr Asn Ser Asp Asn Asp Arg Ser SerArg Leu Leu Leu Glu Arg 685 690 695 ccc tct cca atc aga gac gga cga ggtagt ttg gag aag agc cag ggt 2225 Pro Ser Pro Ile Arg Asp Gly Arg Gly SerLeu Glu Lys Ser Gln Gly 700 705 710 gac aag cga gac cgt aaa aac tct gcatca gct gaa cga gat agg aag 2273 Asp Lys Arg Asp Arg Lys Asn Ser Ala SerAla Glu Arg Asp Arg Lys 715 720 725 730 cac cgg aca act gct ccc act gaggga aaa agc cct ctg aaa aaa gaa 2321 His Arg Thr Thr Ala Pro Thr Glu GlyLys Ser Pro Leu Lys Lys Glu 735 740 745 gac cgc tct gat ggg agt gca cctagc acc agc act gct tcc tcc aag 2369 Asp Arg Ser Asp Gly Ser Ala Pro SerThr Ser Thr Ala Ser Ser Lys 750 755 760 ctg aag tcc ccg tcc cag aaa caggat ggg ggg aca gcc cct gtg gca 2417 Leu Lys Ser Pro Ser Gln Lys Gln AspGly Gly Thr Ala Pro Val Ala 765 770 775 tca gcc tct ccc aaa ctc tgt ttggcc tgg cag ggc atg ctt cta ctg 2465 Ser Ala Ser Pro Lys Leu Cys Leu AlaTrp Gln Gly Met Leu Leu Leu 780 785 790 aag aac agc aac ttt cct tcc aacatg cat ctg ttg cag ggt gac ctc 2513 Lys Asn Ser Asn Phe Pro Ser Asn MetHis Leu Leu Gln Gly Asp Leu 795 800 805 810 caa gtg gct agt agt ctt cttgtg gag ggt tca act gga ggc aaa gtg 2561 Gln Val Ala Ser Ser Leu Leu ValGlu Gly Ser Thr Gly Gly Lys Val 815 820 825 gcc cag ctc aag atc act cagcgt ctc cgt ttg gac cag ccc aag ttg 2609 Ala Gln Leu Lys Ile Thr Gln ArgLeu Arg Leu Asp Gln Pro Lys Leu 830 835 840 gat gaa gta act cga cgc atcaaa gta gca ggg ccc aat ggt tat gcc 2657 Asp Glu Val Thr Arg Arg Ile LysVal Ala Gly Pro Asn Gly Tyr Ala 845 850 855 att ctt ttg gct gtg cct ggaagt tct gac agc cgg tcc tcc tct tcc 2705 Ile Leu Leu Ala Val Pro Gly SerSer Asp Ser Arg Ser Ser Ser Ser 860 865 870 tca gct gca tca gac act gccact tct act cag agg cca ctt agg aac 2753 Ser Ala Ala Ser Asp Thr Ala ThrSer Thr Gln Arg Pro Leu Arg Asn 875 880 885 890 ctt gtg tcc tat tta aagcaa aag cag gca gcc ggg gtg atc agc ctc 2801 Leu Val Ser Tyr Leu Lys GlnLys Gln Ala Ala Gly Val Ile Ser Leu 895 900 905 cct gtg ggg ggc aac aaagac aag gaa aac acc ggg gtc ctt cat gcc 2849 Pro Val Gly Gly Asn Lys AspLys Glu Asn Thr Gly Val Leu His Ala 910 915 920 ttc cca cct tgt gag ttctcc cag cag ttc ctg gat tcc cct gcc aag 2897 Phe Pro Pro Cys Glu Phe SerGln Gln Phe Leu Asp Ser Pro Ala Lys 925 930 935 gca ctg gcc aaa tct gaagaa gat tac ctg gtc atg atc att gtc cgt 2945 Ala Leu Ala Lys Ser Glu GluAsp Tyr Leu Val Met Ile Ile Val Arg 940 945 950 gca aaa ctg gtt gaa cagcgg atg aag ata tgg aat tca aag ctc 2990 Ala Lys Leu Val Glu Gln Arg MetLys Ile Trp Asn Ser Lys Leu 955 960 965 taatggacct ttttgaagag aagttgtggcttatgtggag tttacatggg cctctgatgg 3050 aagaaagcta atctgtttag tatttgtgcattttactaaa atggcagctt aaagttgtgt 3110 atctgctatt gtgatgccaa tgccggtgttttaagtggaa aaaaaatgac ctctttgatt 3170 tgtgctgtgt acacaagatt tctggaaaagtaaagaaaaa ccctttttat ggctcacaca 3230 gcttaagagt agctgtctct caaacgtgcgctcacagttg agctgctttt gttttattct 3290 aaataaattg tttcttttga gg 3312 8969 PRT Homo sapiens 8 Met Arg Thr Ala Gly Arg Asp Pro Val Pro Arg ArgSer Pro Arg Trp 1 5 10 15 Arg Arg Ala Val Pro Leu Cys Glu Thr Ser AlaGly Arg Arg Val Thr 20 25 30 Gln Leu Arg Gly Asp Asp Leu Arg Arg Pro AlaThr Met Lys Gly Lys 35 40 45 Glu Arg Ser Pro Val Lys Ala Lys Arg Ser ArgGly Gly Glu Asp Ser 50 55 60 Thr Ser Arg Gly Glu Arg Ser Lys Lys Leu GlyGly Ser Gly Gly Ser 65 70 75 80 Asn Gly Ser Ser Ser Gly Lys Thr Asp SerGly Gly Gly Ser Arg Arg 85 90 95 Ser Leu Leu Leu Asp Lys Ser Ser Ser ArgGly Gly Ser Arg Glu Tyr 100 105 110 Asp Thr Gly Gly Gly Ser Ser Ser SerArg Leu His Ser Tyr Ser Ser 115 120 125 Pro Ser Thr Lys Asn Ser Ser GlyGly Gly Glu Ser Arg Ser Ser Ser 130 135 140 Arg Gly Gly Gly Gly Glu SerArg Ser Ser Gly Ala Ala Ser Ser Ala 145 150 155 160 Pro Gly Gly Gly AspGly Ala Glu Tyr Lys Thr Leu Lys Ile Ser Glu 165 170 175 Leu Gly Ser GlnLeu Ser Asp Glu Ala Val Glu Asp Gly Leu Phe His 180 185 190 Glu Phe LysArg Phe Gly Asp Val Ser Val Lys Ile Ser His Leu Ser 195 200 205 Gly SerGly Ser Gly Asp Glu Arg Val Ala Phe Val Asn Phe Arg Arg 210 215 220 ProGlu Asp Ala Arg Ala Ala Lys His Ala Arg Gly Arg Leu Val Leu 225 230 235240 Tyr Asp Arg Pro Leu Lys Ile Glu Ala Val Tyr Val Ser Arg Arg Arg 245250 255 Ser Arg Ser Pro Leu Asp Lys Asp Thr Tyr Pro Pro Ser Ala Ser Val260 265 270 Val Gly Ala Ser Val Gly Gly His Arg His Pro Pro Gly Gly GlyGly 275 280 285 Gly Gln Arg Ser Leu Ser Pro Gly Gly Ala Ala Leu Gly TyrArg Asp 290 295 300 Tyr Arg Leu Gln Gln Leu Ala Leu Gly Arg Leu Pro ProPro Pro Pro 305 310 315 320 Pro Pro Leu Pro Arg Asp Leu Glu Arg Glu ArgAsp Tyr Pro Phe Tyr 325 330 335 Glu Arg Val Arg Pro Ala Tyr Ser Leu GluPro Arg Val Gly Ala Gly 340 345 350 Ala Gly Ala Ala Pro Phe Arg Glu ValAsp Glu Ile Ser Pro Glu Asp 355 360 365 Asp Gln Arg Ala Asn Arg Thr LeuPhe Leu Gly Asn Leu Asp Ile Thr 370 375 380 Val Thr Glu Ser Asp Leu ArgArg Ala Phe Asp Arg Phe Gly Val Ile 385 390 395 400 Thr Glu Val Asp IleLys Arg Pro Ser Arg Gly Gln Thr Ser Thr Tyr 405 410 415 Gly Phe Leu LysPhe Glu Asn Leu Asp Met Ser His Arg Ala Lys Leu 420 425 430 Ala Met SerGly Lys Ile Ile Ile Arg Asn Pro Ile Lys Ile Gly Tyr 435 440 445 Gly LysAla Thr Pro Thr Thr Arg Leu Trp Val Gly Gly Leu Gly Pro 450 455 460 TrpVal Pro Leu Ala Ala Leu Ala Arg Glu Phe Asp Arg Phe Gly Thr 465 470 475480 Ile Arg Thr Ile Asp Tyr Arg Lys Gly Asp Ser Trp Ala Tyr Ile Gln 485490 495 Tyr Glu Ser Leu Asp Ala Ala His Ala Ala Trp Thr His Met Arg Gly500 505 510 Phe Pro Leu Gly Gly Pro Asp Arg Arg Leu Arg Val Asp Phe AlaAsp 515 520 525 Thr Glu His Arg Tyr Gln Gln Gln Tyr Leu Gln Pro Leu ProLeu Thr 530 535 540 His Tyr Glu Leu Val Thr Asp Ala Phe Gly His Arg AlaPro Asp Pro 545 550 555 560 Leu Arg Gly Ala Arg Asp Arg Thr Pro Pro LeuLeu Tyr Arg Asp Arg 565 570 575 Asp Arg Asp Leu Tyr Pro Asp Ser Asp TrpVal Pro Pro Pro Pro Pro 580 585 590 Val Arg Glu Arg Ser Thr Arg Thr AlaAla Thr Ser Val Pro Ala Tyr 595 600 605 Glu Pro Leu Asp Ser Leu Asp ArgArg Arg Asp Gly Trp Ser Leu Asp 610 615 620 Arg Asp Arg Gly Asp Arg AspLeu Pro Ser Ser Arg Asp Gln Pro Arg 625 630 635 640 Lys Arg Arg Leu ProGlu Glu Ser Gly Gly Arg His Leu Asp Arg Ser 645 650 655 Pro Glu Ser AspArg Pro Arg Lys Arg His Cys Ala Pro Ser Pro Asp 660 665 670 Arg Ser ProGlu Leu Ser Ser Ser Arg Asp Arg Tyr Asn Ser Asp Asn 675 680 685 Asp ArgSer Ser Arg Leu Leu Leu Glu Arg Pro Ser Pro Ile Arg Asp 690 695 700 GlyArg Gly Ser Leu Glu Lys Ser Gln Gly Asp Lys Arg Asp Arg Lys 705 710 715720 Asn Ser Ala Ser Ala Glu Arg Asp Arg Lys His Arg Thr Thr Ala Pro 725730 735 Thr Glu Gly Lys Ser Pro Leu Lys Lys Glu Asp Arg Ser Asp Gly Ser740 745 750 Ala Pro Ser Thr Ser Thr Ala Ser Ser Lys Leu Lys Ser Pro SerGln 755 760 765 Lys Gln Asp Gly Gly Thr Ala Pro Val Ala Ser Ala Ser ProLys Leu 770 775 780 Cys Leu Ala Trp Gln Gly Met Leu Leu Leu Lys Asn SerAsn Phe Pro 785 790 795 800 Ser Asn Met His Leu Leu Gln Gly Asp Leu GlnVal Ala Ser Ser Leu 805 810 815 Leu Val Glu Gly Ser Thr Gly Gly Lys ValAla Gln Leu Lys Ile Thr 820 825 830 Gln Arg Leu Arg Leu Asp Gln Pro LysLeu Asp Glu Val Thr Arg Arg 835 840 845 Ile Lys Val Ala Gly Pro Asn GlyTyr Ala Ile Leu Leu Ala Val Pro 850 855 860 Gly Ser Ser Asp Ser Arg SerSer Ser Ser Ser Ala Ala Ser Asp Thr 865 870 875 880 Ala Thr Ser Thr GlnArg Pro Leu Arg Asn Leu Val Ser Tyr Leu Lys 885 890 895 Gln Lys Gln AlaAla Gly Val Ile Ser Leu Pro Val Gly Gly Asn Lys 900 905 910 Asp Lys GluAsn Thr Gly Val Leu His Ala Phe Pro Pro Cys Glu Phe 915 920 925 Ser GlnGln Phe Leu Asp Ser Pro Ala Lys Ala Leu Ala Lys Ser Glu 930 935 940 GluAsp Tyr Leu Val Met Ile Ile Val Arg Ala Lys Leu Val Glu Gln 945 950 955960 Arg Met Lys Ile Trp Asn Ser Lys Leu 965 9 3423 DNA Homo sapiens CDS(84)..(3014) 9 ggcgccttcc tggagcgcgg ggagatgtaa agatagacaa ataattttcccaatgagact 60 gtagaagaga gagcaattgg cca atg agg act gcg ggg cgg gac cctgtg ccg 113 Met Arg Thr Ala Gly Arg Asp Pro Val Pro 1 5 10 cgg cgg agtcca aga tgg cgg cgt gcg gtt ccg ctg tgt gaa acg agc 161 Arg Arg Ser ProArg Trp Arg Arg Ala Val Pro Leu Cys Glu Thr Ser 15 20 25 gcg ggg cgg cgggtt act cag ctc cgc gga gac gac ctc cga cga ccc 209 Ala Gly Arg Arg ValThr Gln Leu Arg Gly Asp Asp Leu Arg Arg Pro 30 35 40 gca aca atg aag ggaaaa gag cgc tcg cca gtg aag gcc aaa cgc tcc 257 Ala Thr Met Lys Gly LysGlu Arg Ser Pro Val Lys Ala Lys Arg Ser 45 50 55 cgt ggt ggt gag gac tcgact tcc cgc ggt gag cgg agc aag aag tta 305 Arg Gly Gly Glu Asp Ser ThrSer Arg Gly Glu Arg Ser Lys Lys Leu 60 65 70 ggg ggc tct ggt ggc agc aatggg agc agc agc gga aag acc gat agc 353 Gly Gly Ser Gly Gly Ser Asn GlySer Ser Ser Gly Lys Thr Asp Ser 75 80 85 90 ggc ggt ggg tcg cgg cgg agtctc ctc ctg gac aag tcc agc agt cga 401 Gly Gly Gly Ser Arg Arg Ser LeuLeu Leu Asp Lys Ser Ser Ser Arg 95 100 105 ggt ggc agc cgc gag tat gatacc ggt ggg ggc agc tcc agt agc cgc 449 Gly Gly Ser Arg Glu Tyr Asp ThrGly Gly Gly Ser Ser Ser Ser Arg 110 115 120 ttg cat agt tat agc tcc ccgagc acc aaa aat tct tcg ggc ggg ggc 497 Leu His Ser Tyr Ser Ser Pro SerThr Lys Asn Ser Ser Gly Gly Gly 125 130 135 gag tcg cgc agc agc tcc cggggt gga ggc ggg gag tca cgt tcc tct 545 Glu Ser Arg Ser Ser Ser Arg GlyGly Gly Gly Glu Ser Arg Ser Ser 140 145 150 ggg gcc gcc tcc tca gct cccggc ggc ggg gac ggc gcg gaa tac aag 593 Gly Ala Ala Ser Ser Ala Pro GlyGly Gly Asp Gly Ala Glu Tyr Lys 155 160 165 170 act ctg aag ata agc gagttg ggg tcc cag ctt agt gac gaa gcg gtg 641 Thr Leu Lys Ile Ser Glu LeuGly Ser Gln Leu Ser Asp Glu Ala Val 175 180 185 gag gac ggc ctg ttt catgag ttc aaa cgc ttc ggt gat gta agt gtg 689 Glu Asp Gly Leu Phe His GluPhe Lys Arg Phe Gly Asp Val Ser Val 190 195 200 aaa atc agt cat ctg tcgggt tct ggc agc ggg gat gag cgg gta gcc 737 Lys Ile Ser His Leu Ser GlySer Gly Ser Gly Asp Glu Arg Val Ala 205 210 215 ttt gtg aac ttc cgg cggcca gag gac gcg cgg gcg gcc aag cat gcc 785 Phe Val Asn Phe Arg Arg ProGlu Asp Ala Arg Ala Ala Lys His Ala 220 225 230 aga ggc cgc ctg gtg ctctat gac cgg cct ctg aag ata gaa gct gtg 833 Arg Gly Arg Leu Val Leu TyrAsp Arg Pro Leu Lys Ile Glu Ala Val 235 240 245 250 tat gtg agc cgg cgccgc agc cgc tcc cct tta gac aaa gat act tat 881 Tyr Val Ser Arg Arg ArgSer Arg Ser Pro Leu Asp Lys Asp Thr Tyr 255 260 265 cct cca tca gcc agtgtg gtc ggg gcc tct gta ggt ggt cac cgg cac 929 Pro Pro Ser Ala Ser ValVal Gly Ala Ser Val Gly Gly His Arg His 270 275 280 ccc cct gga ggt ggtgga ggc cag aga tca ctt tcc cct ggt ggc gct 977 Pro Pro Gly Gly Gly GlyGly Gln Arg Ser Leu Ser Pro Gly Gly Ala 285 290 295 gct ttg gga tac agagac tac cgg ctg cag cag ttg gct ctt ggc cgc 1025 Ala Leu Gly Tyr Arg AspTyr Arg Leu Gln Gln Leu Ala Leu Gly Arg 300 305 310 ctg ccc cct cca cctccg cca cca ttg cct cga gac ctg gag aga gaa 1073 Leu Pro Pro Pro Pro ProPro Pro Leu Pro Arg Asp Leu Glu Arg Glu 315 320 325 330 aga gac tac ccgttc tat gag aga gtg cgc cct gca tac agt ctt gag 1121 Arg Asp Tyr Pro PheTyr Glu Arg Val Arg Pro Ala Tyr Ser Leu Glu 335 340 345 cca agg gtg ggagct gga gca ggt gct gct cct ttc aga gaa gtg gat 1169 Pro Arg Val Gly AlaGly Ala Gly Ala Ala Pro Phe Arg Glu Val Asp 350 355 360 gag att tca cccgag gat gat cag cga gct aac cgg acg ctc ttc ttg 1217 Glu Ile Ser Pro GluAsp Asp Gln Arg Ala Asn Arg Thr Leu Phe Leu 365 370 375 ggc aac cta gacatc act gta acg gag agt gat tta aga agg gcg ttt 1265 Gly Asn Leu Asp IleThr Val Thr Glu Ser Asp Leu Arg Arg Ala Phe 380 385 390 gat cgc ttt ggagtc atc aca gaa gta gat atc aag agg cct tct cgc 1313 Asp Arg Phe Gly ValIle Thr Glu Val Asp Ile Lys Arg Pro Ser Arg 395 400 405 410 ggc cag actagt act tac ggc ttt ctc aaa ttt gag aac tta gat atg 1361 Gly Gln Thr SerThr Tyr Gly Phe Leu Lys Phe Glu Asn Leu Asp Met 415 420 425 tct cac cgggcc aaa tta gca atg tct ggc aaa att ata att cgg aat 1409 Ser His Arg AlaLys Leu Ala Met Ser Gly Lys Ile Ile Ile Arg Asn 430 435 440 cct atc aaaatt ggt tat ggt aaa gct aca ccc acc acc cgc ctc tgg 1457 Pro Ile Lys IleGly Tyr Gly Lys Ala Thr Pro Thr Thr Arg Leu Trp 445 450 455 gtg gga ggcctg gga cct tgg gtt cct ctt gct gcc ctg gca cga gaa 1505 Val Gly Gly LeuGly Pro Trp Val Pro Leu Ala Ala Leu Ala Arg Glu 460 465 470 ttt gat cgattt ggc acc ata cgc acc ata gac tac cga aaa ggt gat 1553 Phe Asp Arg PheGly Thr Ile Arg Thr Ile Asp Tyr Arg Lys Gly Asp 475 480 485 490 agt tgggca tat atc cag tat gaa agc ctg gat gca gcg cat gct gcc 1601 Ser Trp AlaTyr Ile Gln Tyr Glu Ser Leu Asp Ala Ala His Ala Ala 495 500 505 tgg acccat atg cgg ggc ttc cca ctt ggt ggc cca gat cga cgc ctt 1649 Trp Thr HisMet Arg Gly Phe Pro Leu Gly Gly Pro Asp Arg Arg Leu 510 515 520 aga gtagac ttt gcc gac acc gaa cat cgt tac cag cag cag tat ctg 1697 Arg Val AspPhe Ala Asp Thr Glu His Arg Tyr Gln Gln Gln Tyr Leu 525 530 535 cag cctctg ccc ttg act cat tat gag ctg gtg aca gat gct ttt gga 1745 Gln Pro LeuPro Leu Thr His Tyr Glu Leu Val Thr Asp Ala Phe Gly 540 545 550 cat cgggca cca gac cct ttg agg ggt gct cgg gat agg aca cca ccc 1793 His Arg AlaPro Asp Pro Leu Arg Gly Ala Arg Asp Arg Thr Pro Pro 555 560 565 570 ttacta tac aga gat cgt gat agg gac ctt tat cct gac tct gat tgg 1841 Leu LeuTyr Arg Asp Arg Asp Arg Asp Leu Tyr Pro Asp Ser Asp Trp 575 580 585 gtgcca ccc cca ccc cca gtc cga gaa cgc agc act cgg act gca gct 1889 Val ProPro Pro Pro Pro Val Arg Glu Arg Ser Thr Arg Thr Ala Ala 590 595 600 acttct gtg cct gct tat gag cca ctg gat agc cta gat cgc agg cgg 1937 Thr SerVal Pro Ala Tyr Glu Pro Leu Asp Ser Leu Asp Arg Arg Arg 605 610 615 gatggt tgg tcc ttg gac cgg gac aga ggt gat cga gat ctg ccc agc 1985 Asp GlyTrp Ser Leu Asp Arg Asp Arg Gly Asp Arg Asp Leu Pro Ser 620 625 630 agcaga gac cag cct agg aag cga agg ctg cct gag gag agt gga gga 2033 Ser ArgAsp Gln Pro Arg Lys Arg Arg Leu Pro Glu Glu Ser Gly Gly 635 640 645 650cgt cat ctg gat agg tct cct gag agt gac cgc cca cga aaa cgt cac 2081 ArgHis Leu Asp Arg Ser Pro Glu Ser Asp Arg Pro Arg Lys Arg His 655 660 665tgc gct cct tct cct gac cgc agt cca gaa ttg agc agt agc cgg gat 2129 CysAla Pro Ser Pro Asp Arg Ser Pro Glu Leu Ser Ser Ser Arg Asp 670 675 680cgt tac aac agc gac aat gat cga tct tcc cgt ctt ctc ttg gaa agg 2177 ArgTyr Asn Ser Asp Asn Asp Arg Ser Ser Arg Leu Leu Leu Glu Arg 685 690 695ccc tct cca atc aga gac gga cga ggt agt ttg gag aag agc cag ggt 2225 ProSer Pro Ile Arg Asp Gly Arg Gly Ser Leu Glu Lys Ser Gln Gly 700 705 710gac aag cga gac cgt aaa aac tct gca tca gct gaa cga gat agg aag 2273 AspLys Arg Asp Arg Lys Asn Ser Ala Ser Ala Glu Arg Asp Arg Lys 715 720 725730 cac cgg aca act gct ccc act gag gga aaa agc cct ctg aaa aaa gaa 2321His Arg Thr Thr Ala Pro Thr Glu Gly Lys Ser Pro Leu Lys Lys Glu 735 740745 gac cgc tct gat ggg agt gca cct agc acc agc act gct tcc tcc aag 2369Asp Arg Ser Asp Gly Ser Ala Pro Ser Thr Ser Thr Ala Ser Ser Lys 750 755760 ctg aag tcc ccg tcc cag aaa cag gat ggg ggg aca gcc cct gtg gca 2417Leu Lys Ser Pro Ser Gln Lys Gln Asp Gly Gly Thr Ala Pro Val Ala 765 770775 tca gcc tct ccc aaa ctc tgt ttg gcc tgg cag ggc atg ctt cta ctg 2465Ser Ala Ser Pro Lys Leu Cys Leu Ala Trp Gln Gly Met Leu Leu Leu 780 785790 aag aac agc aac ttt cct tcc aac atg cat ctg ttg cag ggt gac ctc 2513Lys Asn Ser Asn Phe Pro Ser Asn Met His Leu Leu Gln Gly Asp Leu 795 800805 810 caa gtg gct agt agt ctt ctt gtg gag ggt tca act gga ggc aaa gtg2561 Gln Val Ala Ser Ser Leu Leu Val Glu Gly Ser Thr Gly Gly Lys Val 815820 825 gcc cag ctc aag atc act cag cgt ctc cgt ttg gac cag ccc aag ttg2609 Ala Gln Leu Lys Ile Thr Gln Arg Leu Arg Leu Asp Gln Pro Lys Leu 830835 840 gat gaa gta act cga cgc atc aaa gta gca ggg ccc aat ggt tat gcc2657 Asp Glu Val Thr Arg Arg Ile Lys Val Ala Gly Pro Asn Gly Tyr Ala 845850 855 att ctt ttg gct gtg cct gga agt tct gac agc cgg tcc tcc tct tcc2705 Ile Leu Leu Ala Val Pro Gly Ser Ser Asp Ser Arg Ser Ser Ser Ser 860865 870 tca gct gca tca gac act gcc act tct act cag agg cca ctt agg aac2753 Ser Ala Ala Ser Asp Thr Ala Thr Ser Thr Gln Arg Pro Leu Arg Asn 875880 885 890 ctt gtg tcc tat tta aag caa aag cag gca gcc ggg gtg atc agcctc 2801 Leu Val Ser Tyr Leu Lys Gln Lys Gln Ala Ala Gly Val Ile Ser Leu895 900 905 cct gtg ggg ggc aac aaa gac aag gaa aac acc ggg gtc ctt catgcc 2849 Pro Val Gly Gly Asn Lys Asp Lys Glu Asn Thr Gly Val Leu His Ala910 915 920 ttc cca cct tgt gag ttc tcc cag cag ttc ctg gat tcc cct gccaag 2897 Phe Pro Pro Cys Glu Phe Ser Gln Gln Phe Leu Asp Ser Pro Ala Lys925 930 935 gca ctg gcc aaa tct gaa gaa gat tac ctg gtc atg atc att gtccgt 2945 Ala Leu Ala Lys Ser Glu Glu Asp Tyr Leu Val Met Ile Ile Val Arg940 945 950 ggg ttt ggt ttt cag ata gga gtt agg tat gag aac aag aag agagaa 2993 Gly Phe Gly Phe Gln Ile Gly Val Arg Tyr Glu Asn Lys Lys Arg Glu955 960 965 970 aac ttg gcg ctg acc ctg tta tagtggttat agtggtgtccctaaagggag 3044 Asn Leu Ala Leu Thr Leu Leu 975 gaaatgattt cagcaaaactggttgaacag cggatgaaga tatggaattc aaagctctaa 3104 tggacctttt tgaagagaagttgtggctta tgtggagttt acatgggcct ctgatggaag 3164 aaagctaatc tgtttagtatttgtgcattt tactaaaatg gcagcttaaa gttgtgtatc 3224 tgctattgtg atgccaatgccggtgtttta agtggaaaaa aaatgacctc tttgatttgt 3284 gctgtgtaca caagatttctggaaaagtaa agaaaaaccc tttttatggc tcacacagct 3344 taagagtagc tgtctctcaaacgtgcgctc acagttgagc tgcttttgtt ttattctaaa 3404 taaattgttt cttttgagg3423 10 977 PRT Homo sapiens 10 Met Arg Thr Ala Gly Arg Asp Pro Val ProArg Arg Ser Pro Arg Trp 1 5 10 15 Arg Arg Ala Val Pro Leu Cys Glu ThrSer Ala Gly Arg Arg Val Thr 20 25 30 Gln Leu Arg Gly Asp Asp Leu Arg ArgPro Ala Thr Met Lys Gly Lys 35 40 45 Glu Arg Ser Pro Val Lys Ala Lys ArgSer Arg Gly Gly Glu Asp Ser 50 55 60 Thr Ser Arg Gly Glu Arg Ser Lys LysLeu Gly Gly Ser Gly Gly Ser 65 70 75 80 Asn Gly Ser Ser Ser Gly Lys ThrAsp Ser Gly Gly Gly Ser Arg Arg 85 90 95 Ser Leu Leu Leu Asp Lys Ser SerSer Arg Gly Gly Ser Arg Glu Tyr 100 105 110 Asp Thr Gly Gly Gly Ser SerSer Ser Arg Leu His Ser Tyr Ser Ser 115 120 125 Pro Ser Thr Lys Asn SerSer Gly Gly Gly Glu Ser Arg Ser Ser Ser 130 135 140 Arg Gly Gly Gly GlyGlu Ser Arg Ser Ser Gly Ala Ala Ser Ser Ala 145 150 155 160 Pro Gly GlyGly Asp Gly Ala Glu Tyr Lys Thr Leu Lys Ile Ser Glu 165 170 175 Leu GlySer Gln Leu Ser Asp Glu Ala Val Glu Asp Gly Leu Phe His 180 185 190 GluPhe Lys Arg Phe Gly Asp Val Ser Val Lys Ile Ser His Leu Ser 195 200 205Gly Ser Gly Ser Gly Asp Glu Arg Val Ala Phe Val Asn Phe Arg Arg 210 215220 Pro Glu Asp Ala Arg Ala Ala Lys His Ala Arg Gly Arg Leu Val Leu 225230 235 240 Tyr Asp Arg Pro Leu Lys Ile Glu Ala Val Tyr Val Ser Arg ArgArg 245 250 255 Ser Arg Ser Pro Leu Asp Lys Asp Thr Tyr Pro Pro Ser AlaSer Val 260 265 270 Val Gly Ala Ser Val Gly Gly His Arg His Pro Pro GlyGly Gly Gly 275 280 285 Gly Gln Arg Ser Leu Ser Pro Gly Gly Ala Ala LeuGly Tyr Arg Asp 290 295 300 Tyr Arg Leu Gln Gln Leu Ala Leu Gly Arg LeuPro Pro Pro Pro Pro 305 310 315 320 Pro Pro Leu Pro Arg Asp Leu Glu ArgGlu Arg Asp Tyr Pro Phe Tyr 325 330 335 Glu Arg Val Arg Pro Ala Tyr SerLeu Glu Pro Arg Val Gly Ala Gly 340 345 350 Ala Gly Ala Ala Pro Phe ArgGlu Val Asp Glu Ile Ser Pro Glu Asp 355 360 365 Asp Gln Arg Ala Asn ArgThr Leu Phe Leu Gly Asn Leu Asp Ile Thr 370 375 380 Val Thr Glu Ser AspLeu Arg Arg Ala Phe Asp Arg Phe Gly Val Ile 385 390 395 400 Thr Glu ValAsp Ile Lys Arg Pro Ser Arg Gly Gln Thr Ser Thr Tyr 405 410 415 Gly PheLeu Lys Phe Glu Asn Leu Asp Met Ser His Arg Ala Lys Leu 420 425 430 AlaMet Ser Gly Lys Ile Ile Ile Arg Asn Pro Ile Lys Ile Gly Tyr 435 440 445Gly Lys Ala Thr Pro Thr Thr Arg Leu Trp Val Gly Gly Leu Gly Pro 450 455460 Trp Val Pro Leu Ala Ala Leu Ala Arg Glu Phe Asp Arg Phe Gly Thr 465470 475 480 Ile Arg Thr Ile Asp Tyr Arg Lys Gly Asp Ser Trp Ala Tyr IleGln 485 490 495 Tyr Glu Ser Leu Asp Ala Ala His Ala Ala Trp Thr His MetArg Gly 500 505 510 Phe Pro Leu Gly Gly Pro Asp Arg Arg Leu Arg Val AspPhe Ala Asp 515 520 525 Thr Glu His Arg Tyr Gln Gln Gln Tyr Leu Gln ProLeu Pro Leu Thr 530 535 540 His Tyr Glu Leu Val Thr Asp Ala Phe Gly HisArg Ala Pro Asp Pro 545 550 555 560 Leu Arg Gly Ala Arg Asp Arg Thr ProPro Leu Leu Tyr Arg Asp Arg 565 570 575 Asp Arg Asp Leu Tyr Pro Asp SerAsp Trp Val Pro Pro Pro Pro Pro 580 585 590 Val Arg Glu Arg Ser Thr ArgThr Ala Ala Thr Ser Val Pro Ala Tyr 595 600 605 Glu Pro Leu Asp Ser LeuAsp Arg Arg Arg Asp Gly Trp Ser Leu Asp 610 615 620 Arg Asp Arg Gly AspArg Asp Leu Pro Ser Ser Arg Asp Gln Pro Arg 625 630 635 640 Lys Arg ArgLeu Pro Glu Glu Ser Gly Gly Arg His Leu Asp Arg Ser 645 650 655 Pro GluSer Asp Arg Pro Arg Lys Arg His Cys Ala Pro Ser Pro Asp 660 665 670 ArgSer Pro Glu Leu Ser Ser Ser Arg Asp Arg Tyr Asn Ser Asp Asn 675 680 685Asp Arg Ser Ser Arg Leu Leu Leu Glu Arg Pro Ser Pro Ile Arg Asp 690 695700 Gly Arg Gly Ser Leu Glu Lys Ser Gln Gly Asp Lys Arg Asp Arg Lys 705710 715 720 Asn Ser Ala Ser Ala Glu Arg Asp Arg Lys His Arg Thr Thr AlaPro 725 730 735 Thr Glu Gly Lys Ser Pro Leu Lys Lys Glu Asp Arg Ser AspGly Ser 740 745 750 Ala Pro Ser Thr Ser Thr Ala Ser Ser Lys Leu Lys SerPro Ser Gln 755 760 765 Lys Gln Asp Gly Gly Thr Ala Pro Val Ala Ser AlaSer Pro Lys Leu 770 775 780 Cys Leu Ala Trp Gln Gly Met Leu Leu Leu LysAsn Ser Asn Phe Pro 785 790 795 800 Ser Asn Met His Leu Leu Gln Gly AspLeu Gln Val Ala Ser Ser Leu 805 810 815 Leu Val Glu Gly Ser Thr Gly GlyLys Val Ala Gln Leu Lys Ile Thr 820 825 830 Gln Arg Leu Arg Leu Asp GlnPro Lys Leu Asp Glu Val Thr Arg Arg 835 840 845 Ile Lys Val Ala Gly ProAsn Gly Tyr Ala Ile Leu Leu Ala Val Pro 850 855 860 Gly Ser Ser Asp SerArg Ser Ser Ser Ser Ser Ala Ala Ser Asp Thr 865 870 875 880 Ala Thr SerThr Gln Arg Pro Leu Arg Asn Leu Val Ser Tyr Leu Lys 885 890 895 Gln LysGln Ala Ala Gly Val Ile Ser Leu Pro Val Gly Gly Asn Lys 900 905 910 AspLys Glu Asn Thr Gly Val Leu His Ala Phe Pro Pro Cys Glu Phe 915 920 925Ser Gln Gln Phe Leu Asp Ser Pro Ala Lys Ala Leu Ala Lys Ser Glu 930 935940 Glu Asp Tyr Leu Val Met Ile Ile Val Arg Gly Phe Gly Phe Gln Ile 945950 955 960 Gly Val Arg Tyr Glu Asn Lys Lys Arg Glu Asn Leu Ala Leu ThrLeu 965 970 975 Leu 11 3383 DNA Homo sapiens CDS (84)..(2954) 11ggcgccttcc tggagcgcgg ggagatgtaa agatagacaa ataattttcc caatgagact 60gtagaagaga gagcaattgg cca atg agg act gcg ggg cgg gac cct gtg ccg 113Met Arg Thr Ala Gly Arg Asp Pro Val Pro 1 5 10 cgg cgg agt cca aga tggcgg cgt gcg gtt ccg ctg tgt gaa acg agc 161 Arg Arg Ser Pro Arg Trp ArgArg Ala Val Pro Leu Cys Glu Thr Ser 15 20 25 gcg ggg cgg cgg gtt act cagctc cgc gga gac gac ctc cga cga ccc 209 Ala Gly Arg Arg Val Thr Gln LeuArg Gly Asp Asp Leu Arg Arg Pro 30 35 40 gca aca atg aag gga aaa gag cgctcg cca gtg aag gcc aaa cgc tcc 257 Ala Thr Met Lys Gly Lys Glu Arg SerPro Val Lys Ala Lys Arg Ser 45 50 55 cgt ggt ggt gag gac tcg act tcc cgcggt gag cgg agc aag aag tta 305 Arg Gly Gly Glu Asp Ser Thr Ser Arg GlyGlu Arg Ser Lys Lys Leu 60 65 70 ggg ggc tct ggt ggc agc aat ggg agc agcagc gga aag acc gat agc 353 Gly Gly Ser Gly Gly Ser Asn Gly Ser Ser SerGly Lys Thr Asp Ser 75 80 85 90 ggc ggt ggg tcg cgg cgg agt ctc ctc ctggac aag tcc agc agt cga 401 Gly Gly Gly Ser Arg Arg Ser Leu Leu Leu AspLys Ser Ser Ser Arg 95 100 105 ggt ggc agc cgc gag tat gat acc ggt gggggc agc tcc agt agc cgc 449 Gly Gly Ser Arg Glu Tyr Asp Thr Gly Gly GlySer Ser Ser Ser Arg 110 115 120 ttg cat agt tat agc tcc ccg agc acc aaaaat tct tcg ggc ggg ggc 497 Leu His Ser Tyr Ser Ser Pro Ser Thr Lys AsnSer Ser Gly Gly Gly 125 130 135 gag tcg cgc agc agc tcc cgg ggt gga ggcggg gag tca cgt tcc tct 545 Glu Ser Arg Ser Ser Ser Arg Gly Gly Gly GlyGlu Ser Arg Ser Ser 140 145 150 ggg gcc gcc tcc tca gct ccc ggc ggc ggggac ggc gcg gaa tac aag 593 Gly Ala Ala Ser Ser Ala Pro Gly Gly Gly AspGly Ala Glu Tyr Lys 155 160 165 170 act ctg aag ata agc gag ttg ggg tcccag ctt agt gac gaa gcg gtg 641 Thr Leu Lys Ile Ser Glu Leu Gly Ser GlnLeu Ser Asp Glu Ala Val 175 180 185 gag gac ggc ctg ttt cat gag ttc aaacgc ttc ggt gat gta agt gtg 689 Glu Asp Gly Leu Phe His Glu Phe Lys ArgPhe Gly Asp Val Ser Val 190 195 200 aaa atc agt cat ctg tcg ggt tct ggcagc ggg gat gag cgg gta gcc 737 Lys Ile Ser His Leu Ser Gly Ser Gly SerGly Asp Glu Arg Val Ala 205 210 215 ttt gtg aac ttc cgg cgg cca gag gacgcg cgg gcg gcc aag cat gcc 785 Phe Val Asn Phe Arg Arg Pro Glu Asp AlaArg Ala Ala Lys His Ala 220 225 230 aga ggc cgc ctg gtg ctc tat gac cggcct ctg aag ata gaa gct gtg 833 Arg Gly Arg Leu Val Leu Tyr Asp Arg ProLeu Lys Ile Glu Ala Val 235 240 245 250 tat gtg agc cgg cgc cgc agc cgctcc cct tta gac aaa gat act tat 881 Tyr Val Ser Arg Arg Arg Ser Arg SerPro Leu Asp Lys Asp Thr Tyr 255 260 265 cct cca tca gcc agt gtg gtc ggggcc tct gta ggt ggt cac cgg cac 929 Pro Pro Ser Ala Ser Val Val Gly AlaSer Val Gly Gly His Arg His 270 275 280 ccc cct gga ggt ggt gga ggc cagaga tca ctt tcc cct ggt ggc gct 977 Pro Pro Gly Gly Gly Gly Gly Gln ArgSer Leu Ser Pro Gly Gly Ala 285 290 295 gct ttg gga tac aga gac tac cggctg cag cag ttg gct ctt ggc cgc 1025 Ala Leu Gly Tyr Arg Asp Tyr Arg LeuGln Gln Leu Ala Leu Gly Arg 300 305 310 ctg ccc cct cca cct ccg cca ccattg cct cga gac ctg gag aga gaa 1073 Leu Pro Pro Pro Pro Pro Pro Pro LeuPro Arg Asp Leu Glu Arg Glu 315 320 325 330 aga gac tac ccg ttc tat gagaga gtg cgc cct gca tac agt ctt gag 1121 Arg Asp Tyr Pro Phe Tyr Glu ArgVal Arg Pro Ala Tyr Ser Leu Glu 335 340 345 cca agg gtg gga gct gga gcaggt gct gct cct ttc aga gaa gtg gat 1169 Pro Arg Val Gly Ala Gly Ala GlyAla Ala Pro Phe Arg Glu Val Asp 350 355 360 gag att tca ccc gag gat gatcag cga gct aac cgg acg ctc ttc ttg 1217 Glu Ile Ser Pro Glu Asp Asp GlnArg Ala Asn Arg Thr Leu Phe Leu 365 370 375 ggc aac cta gac atc act gtaacg gag agt gat tta aga agg gcg ttt 1265 Gly Asn Leu Asp Ile Thr Val ThrGlu Ser Asp Leu Arg Arg Ala Phe 380 385 390 gat cgc ttt gga gtc atc acagaa gta gat atc aag agg cct tct cgc 1313 Asp Arg Phe Gly Val Ile Thr GluVal Asp Ile Lys Arg Pro Ser Arg 395 400 405 410 ggc cag act agt act tacggc ttt ctc aaa ttt gag aac tta gat atg 1361 Gly Gln Thr Ser Thr Tyr GlyPhe Leu Lys Phe Glu Asn Leu Asp Met 415 420 425 tct cac cgg gcc aaa ttagca atg tct ggc aaa att ata att cgg aat 1409 Ser His Arg Ala Lys Leu AlaMet Ser Gly Lys Ile Ile Ile Arg Asn 430 435 440 cct atc aaa att ggt tatggt aaa gct aca ccc acc acc cgc ctc tgg 1457 Pro Ile Lys Ile Gly Tyr GlyLys Ala Thr Pro Thr Thr Arg Leu Trp 445 450 455 gtg gga ggc ctg gga ccttgg gtt cct ctt gct gcc ctg gca cga gaa 1505 Val Gly Gly Leu Gly Pro TrpVal Pro Leu Ala Ala Leu Ala Arg Glu 460 465 470 ttt gat cga ttt ggc accata cgc acc ata gac tac cga aaa ggt gat 1553 Phe Asp Arg Phe Gly Thr IleArg Thr Ile Asp Tyr Arg Lys Gly Asp 475 480 485 490 agt tgg gca tat atccag tat gaa agc ctg gat gca gcg cat gct gcc 1601 Ser Trp Ala Tyr Ile GlnTyr Glu Ser Leu Asp Ala Ala His Ala Ala 495 500 505 tgg acc cat atg cggggc ttc cca ctt ggt ggc cca gat cga cgc ctt 1649 Trp Thr His Met Arg GlyPhe Pro Leu Gly Gly Pro Asp Arg Arg Leu 510 515 520 aga gta gac ttt gccgac acc gaa cat cgt tac cag cag cag tat ctg 1697 Arg Val Asp Phe Ala AspThr Glu His Arg Tyr Gln Gln Gln Tyr Leu 525 530 535 cag cct ctg ccc ttgact cat tat gag ctg gtg aca gat gct ttt gga 1745 Gln Pro Leu Pro Leu ThrHis Tyr Glu Leu Val Thr Asp Ala Phe Gly 540 545 550 cat cgg gca cca gaccct ttg agg ggt gct cgg gat agg aca cca ccc 1793 His Arg Ala Pro Asp ProLeu Arg Gly Ala Arg Asp Arg Thr Pro Pro 555 560 565 570 tta cta tac agagat cgt gat agg gac ctt tat cct gac tct gat tgg 1841 Leu Leu Tyr Arg AspArg Asp Arg Asp Leu Tyr Pro Asp Ser Asp Trp 575 580 585 gtg cca ccc ccaccc cca gtc cga gaa cgc agc act cgg act gca gct 1889 Val Pro Pro Pro ProPro Val Arg Glu Arg Ser Thr Arg Thr Ala Ala 590 595 600 act tct gtg cctgct tat gag cca ctg gat agc cta gat cgc agg cgg 1937 Thr Ser Val Pro AlaTyr Glu Pro Leu Asp Ser Leu Asp Arg Arg Arg 605 610 615 gat ggt tgg tccttg gac cgg gac aga ggt gat cga gat ctg ccc agc 1985 Asp Gly Trp Ser LeuAsp Arg Asp Arg Gly Asp Arg Asp Leu Pro Ser 620 625 630 agc aga gac cagcct agg aag cga agg ctg cct gag gag agt gga gga 2033 Ser Arg Asp Gln ProArg Lys Arg Arg Leu Pro Glu Glu Ser Gly Gly 635 640 645 650 cgt cat ctggat agg tct cct gag agt gac cgc cca cga aaa cgt cac 2081 Arg His Leu AspArg Ser Pro Glu Ser Asp Arg Pro Arg Lys Arg His 655 660 665 tgc gct ccttct cct gac cgc agt cca gaa ttg agc agt agc cgg gat 2129 Cys Ala Pro SerPro Asp Arg Ser Pro Glu Leu Ser Ser Ser Arg Asp 670 675 680 cgt tac aacagc gac aat gat cga tct tcc cgt ctt ctc ttg gaa agg 2177 Arg Tyr Asn SerAsp Asn Asp Arg Ser Ser Arg Leu Leu Leu Glu Arg 685 690 695 ccc tct ccaatc aga gac gga cga ggt agt ttg gag aag agc cag ggt 2225 Pro Ser Pro IleArg Asp Gly Arg Gly Ser Leu Glu Lys Ser Gln Gly 700 705 710 gac aag cgagac cgt aaa aac tct gca tca gct gaa cga gat agg aag 2273 Asp Lys Arg AspArg Lys Asn Ser Ala Ser Ala Glu Arg Asp Arg Lys 715 720 725 730 cac cggaca act gct ccc act gag gga aaa agc cct ctg aaa aaa gaa 2321 His Arg ThrThr Ala Pro Thr Glu Gly Lys Ser Pro Leu Lys Lys Glu 735 740 745 gac cgctct gat ggg agt gca cct agc acc agc act gct tcc tcc aag 2369 Asp Arg SerAsp Gly Ser Ala Pro Ser Thr Ser Thr Ala Ser Ser Lys 750 755 760 ctg aagtcc ccg tcc cag aaa cag gat ggg ggg aca gcc cct gtg gca 2417 Leu Lys SerPro Ser Gln Lys Gln Asp Gly Gly Thr Ala Pro Val Ala 765 770 775 tca gcctct ccc aaa ctc tgt ttg gcc tgg cag ggc atg ctt cta ctg 2465 Ser Ala SerPro Lys Leu Cys Leu Ala Trp Gln Gly Met Leu Leu Leu 780 785 790 aag aacagc aac ttt cct tcc aac atg cat ctg ttg cag ggt gac ctc 2513 Lys Asn SerAsn Phe Pro Ser Asn Met His Leu Leu Gln Gly Asp Leu 795 800 805 810 caagtg gct agt agt ctt ctt gtg gag ggt tca act gga ggc aaa gtg 2561 Gln ValAla Ser Ser Leu Leu Val Glu Gly Ser Thr Gly Gly Lys Val 815 820 825 gcccag ctc aag atc act cag cgt ctc cgt ttg gac cag ccc aag ttg 2609 Ala GlnLeu Lys Ile Thr Gln Arg Leu Arg Leu Asp Gln Pro Lys Leu 830 835 840 gatgaa gta act cga cgc atc aaa gta gca ggg ccc aat ggt tat gcc 2657 Asp GluVal Thr Arg Arg Ile Lys Val Ala Gly Pro Asn Gly Tyr Ala 845 850 855 attctt ttg gct gtg cct gga agt tct gac agc cgg tcc tcc tct tcc 2705 Ile LeuLeu Ala Val Pro Gly Ser Ser Asp Ser Arg Ser Ser Ser Ser 860 865 870 tcagct gca tca gac act gcc act tct act cag agg cca ctt agg aac 2753 Ser AlaAla Ser Asp Thr Ala Thr Ser Thr Gln Arg Pro Leu Arg Asn 875 880 885 890ctt gtg tcc tat tta aag caa aag cag gca gcc ggg gtg atc agc ctc 2801 LeuVal Ser Tyr Leu Lys Gln Lys Gln Ala Ala Gly Val Ile Ser Leu 895 900 905cct gtg ggg ggc aac aaa gac aag gaa aac acc ggg gtc ctt cat gcc 2849 ProVal Gly Gly Asn Lys Asp Lys Glu Asn Thr Gly Val Leu His Ala 910 915 920ttc cca cct tgt gag ttc tcc cag cag ttc ctg gat tcc cct gcc aag 2897 PhePro Pro Cys Glu Phe Ser Gln Gln Phe Leu Asp Ser Pro Ala Lys 925 930 935gca ctg gcc aaa tct gaa gaa gat tac ctg gtc atg atc att gtc cgt 2945 AlaLeu Ala Lys Ser Glu Glu Asp Tyr Leu Val Met Ile Ile Val Arg 940 945 950ggt gcg tcc taaagtccgt gtgtaacttg tatttactac tttgacatgg 2994 Gly Ala Ser955 ttcctgtttt gtgatgtgta atggatacag catcagatgc aattttcttt ttagttgtta3054 gttgtagcat tttctttttt atatttttat aaacgtcttt aaaatagaaa tcaggacagt3114 ttagctattt ttttgtttgt ttagctatta ttttaagtga aagggatgcc ctaaaggtag3174 caggcaggca gacagatttg ctttaattag gagttcccac ccttatgagt aatttttttt3234 ctctattcag ttgttttttt tttaatcttg agcttaaaaa atcctcagag ttacaaaacc3294 aaaattttga aaagtcagaa tttggagaaa ggagtccact gaccatataa agagagtaat3354 agccacctaa aaaaaaaaaa aaaaaaaaa 3383 12 957 PRT Homo sapiens 12 MetArg Thr Ala Gly Arg Asp Pro Val Pro Arg Arg Ser Pro Arg Trp 1 5 10 15Arg Arg Ala Val Pro Leu Cys Glu Thr Ser Ala Gly Arg Arg Val Thr 20 25 30Gln Leu Arg Gly Asp Asp Leu Arg Arg Pro Ala Thr Met Lys Gly Lys 35 40 45Glu Arg Ser Pro Val Lys Ala Lys Arg Ser Arg Gly Gly Glu Asp Ser 50 55 60Thr Ser Arg Gly Glu Arg Ser Lys Lys Leu Gly Gly Ser Gly Gly Ser 65 70 7580 Asn Gly Ser Ser Ser Gly Lys Thr Asp Ser Gly Gly Gly Ser Arg Arg 85 9095 Ser Leu Leu Leu Asp Lys Ser Ser Ser Arg Gly Gly Ser Arg Glu Tyr 100105 110 Asp Thr Gly Gly Gly Ser Ser Ser Ser Arg Leu His Ser Tyr Ser Ser115 120 125 Pro Ser Thr Lys Asn Ser Ser Gly Gly Gly Glu Ser Arg Ser SerSer 130 135 140 Arg Gly Gly Gly Gly Glu Ser Arg Ser Ser Gly Ala Ala SerSer Ala 145 150 155 160 Pro Gly Gly Gly Asp Gly Ala Glu Tyr Lys Thr LeuLys Ile Ser Glu 165 170 175 Leu Gly Ser Gln Leu Ser Asp Glu Ala Val GluAsp Gly Leu Phe His 180 185 190 Glu Phe Lys Arg Phe Gly Asp Val Ser ValLys Ile Ser His Leu Ser 195 200 205 Gly Ser Gly Ser Gly Asp Glu Arg ValAla Phe Val Asn Phe Arg Arg 210 215 220 Pro Glu Asp Ala Arg Ala Ala LysHis Ala Arg Gly Arg Leu Val Leu 225 230 235 240 Tyr Asp Arg Pro Leu LysIle Glu Ala Val Tyr Val Ser Arg Arg Arg 245 250 255 Ser Arg Ser Pro LeuAsp Lys Asp Thr Tyr Pro Pro Ser Ala Ser Val 260 265 270 Val Gly Ala SerVal Gly Gly His Arg His Pro Pro Gly Gly Gly Gly 275 280 285 Gly Gln ArgSer Leu Ser Pro Gly Gly Ala Ala Leu Gly Tyr Arg Asp 290 295 300 Tyr ArgLeu Gln Gln Leu Ala Leu Gly Arg Leu Pro Pro Pro Pro Pro 305 310 315 320Pro Pro Leu Pro Arg Asp Leu Glu Arg Glu Arg Asp Tyr Pro Phe Tyr 325 330335 Glu Arg Val Arg Pro Ala Tyr Ser Leu Glu Pro Arg Val Gly Ala Gly 340345 350 Ala Gly Ala Ala Pro Phe Arg Glu Val Asp Glu Ile Ser Pro Glu Asp355 360 365 Asp Gln Arg Ala Asn Arg Thr Leu Phe Leu Gly Asn Leu Asp IleThr 370 375 380 Val Thr Glu Ser Asp Leu Arg Arg Ala Phe Asp Arg Phe GlyVal Ile 385 390 395 400 Thr Glu Val Asp Ile Lys Arg Pro Ser Arg Gly GlnThr Ser Thr Tyr 405 410 415 Gly Phe Leu Lys Phe Glu Asn Leu Asp Met SerHis Arg Ala Lys Leu 420 425 430 Ala Met Ser Gly Lys Ile Ile Ile Arg AsnPro Ile Lys Ile Gly Tyr 435 440 445 Gly Lys Ala Thr Pro Thr Thr Arg LeuTrp Val Gly Gly Leu Gly Pro 450 455 460 Trp Val Pro Leu Ala Ala Leu AlaArg Glu Phe Asp Arg Phe Gly Thr 465 470 475 480 Ile Arg Thr Ile Asp TyrArg Lys Gly Asp Ser Trp Ala Tyr Ile Gln 485 490 495 Tyr Glu Ser Leu AspAla Ala His Ala Ala Trp Thr His Met Arg Gly 500 505 510 Phe Pro Leu GlyGly Pro Asp Arg Arg Leu Arg Val Asp Phe Ala Asp 515 520 525 Thr Glu HisArg Tyr Gln Gln Gln Tyr Leu Gln Pro Leu Pro Leu Thr 530 535 540 His TyrGlu Leu Val Thr Asp Ala Phe Gly His Arg Ala Pro Asp Pro 545 550 555 560Leu Arg Gly Ala Arg Asp Arg Thr Pro Pro Leu Leu Tyr Arg Asp Arg 565 570575 Asp Arg Asp Leu Tyr Pro Asp Ser Asp Trp Val Pro Pro Pro Pro Pro 580585 590 Val Arg Glu Arg Ser Thr Arg Thr Ala Ala Thr Ser Val Pro Ala Tyr595 600 605 Glu Pro Leu Asp Ser Leu Asp Arg Arg Arg Asp Gly Trp Ser LeuAsp 610 615 620 Arg Asp Arg Gly Asp Arg Asp Leu Pro Ser Ser Arg Asp GlnPro Arg 625 630 635 640 Lys Arg Arg Leu Pro Glu Glu Ser Gly Gly Arg HisLeu Asp Arg Ser 645 650 655 Pro Glu Ser Asp Arg Pro Arg Lys Arg His CysAla Pro Ser Pro Asp 660 665 670 Arg Ser Pro Glu Leu Ser Ser Ser Arg AspArg Tyr Asn Ser Asp Asn 675 680 685 Asp Arg Ser Ser Arg Leu Leu Leu GluArg Pro Ser Pro Ile Arg Asp 690 695 700 Gly Arg Gly Ser Leu Glu Lys SerGln Gly Asp Lys Arg Asp Arg Lys 705 710 715 720 Asn Ser Ala Ser Ala GluArg Asp Arg Lys His Arg Thr Thr Ala Pro 725 730 735 Thr Glu Gly Lys SerPro Leu Lys Lys Glu Asp Arg Ser Asp Gly Ser 740 745 750 Ala Pro Ser ThrSer Thr Ala Ser Ser Lys Leu Lys Ser Pro Ser Gln 755 760 765 Lys Gln AspGly Gly Thr Ala Pro Val Ala Ser Ala Ser Pro Lys Leu 770 775 780 Cys LeuAla Trp Gln Gly Met Leu Leu Leu Lys Asn Ser Asn Phe Pro 785 790 795 800Ser Asn Met His Leu Leu Gln Gly Asp Leu Gln Val Ala Ser Ser Leu 805 810815 Leu Val Glu Gly Ser Thr Gly Gly Lys Val Ala Gln Leu Lys Ile Thr 820825 830 Gln Arg Leu Arg Leu Asp Gln Pro Lys Leu Asp Glu Val Thr Arg Arg835 840 845 Ile Lys Val Ala Gly Pro Asn Gly Tyr Ala Ile Leu Leu Ala ValPro 850 855 860 Gly Ser Ser Asp Ser Arg Ser Ser Ser Ser Ser Ala Ala SerAsp Thr 865 870 875 880 Ala Thr Ser Thr Gln Arg Pro Leu Arg Asn Leu ValSer Tyr Leu Lys 885 890 895 Gln Lys Gln Ala Ala Gly Val Ile Ser Leu ProVal Gly Gly Asn Lys 900 905 910 Asp Lys Glu Asn Thr Gly Val Leu His AlaPhe Pro Pro Cys Glu Phe 915 920 925 Ser Gln Gln Phe Leu Asp Ser Pro AlaLys Ala Leu Ala Lys Ser Glu 930 935 940 Glu Asp Tyr Leu Val Met Ile IleVal Arg Gly Ala Ser 945 950 955 13 4447 DNA Homo sapiens CDS(551)..(3346) 13 tcccatcttt cccaccctct tggttgccgc tggccacacg ccctccgctggcggcgactt 60 ctcagctccg tgcgcccggg ctggacagtg agcctcgaga ggagacgcgggcggctagag 120 ccggagtggg gcgagccgcg gaacccggcc gggagccgcg cgaggcgtgatcggagggta 180 tggttggcat ggaattgaat ttcatctgtc tgtgggaatt gtaagcaagattgccatcac 240 gaaagccaaa gtggatttct ccagtgtggt gtgcctgccc ccttccgtcattgctgtgaa 300 tgggctggac ggaggagggg ccggcgaaaa tgatgatgaa ccagtgctcgtgtccttatc 360 tgcggcaccc agtccccaga gtgaagctgt tgccaatgaa ctgcaggagctctccttgca 420 gcccgagctg accctaggcc tccaccctgg caggaatccc aatttgcctccacttagtga 480 gcggaagaat gtgctacagt tgaaactcca gcagcgccgg acccgggaagaactggtgag 540 ccaagggatc atg ccg cct ttg aaa agt cca gcc gca ttt catgag cag 589 Met Pro Pro Leu Lys Ser Pro Ala Ala Phe His Glu Gln 1 5 10aga agg agc ttg gag cgg gcc agg aca gag gac tat ctc aaa cgg aag 637 ArgArg Ser Leu Glu Arg Ala Arg Thr Glu Asp Tyr Leu Lys Arg Lys 15 20 25 attcgt tcc cgg ccg gag aga tcg gag ctg gtc agg atg cac att ttg 685 Ile ArgSer Arg Pro Glu Arg Ser Glu Leu Val Arg Met His Ile Leu 30 35 40 45 gaagag acc tcg gct gag cca tcc ctc cag gcc aag cag ctg aag ctg 733 Glu GluThr Ser Ala Glu Pro Ser Leu Gln Ala Lys Gln Leu Lys Leu 50 55 60 aag agagcc aga cta gcc gat gac ctc aat gag aag att gca cag agg 781 Lys Arg AlaArg Leu Ala Asp Asp Leu Asn Glu Lys Ile Ala Gln Arg 65 70 75 cct ggc cccatg gag ctg gtg gag aag aac atc ctt cct gtt gag tcc 829 Pro Gly Pro MetGlu Leu Val Glu Lys Asn Ile Leu Pro Val Glu Ser 80 85 90 agc ctg aag gaagcc atc att gtg ggc cag gtg aac tat ccc aaa gta 877 Ser Leu Lys Glu AlaIle Ile Val Gly Gln Val Asn Tyr Pro Lys Val 95 100 105 gca gac agc tcttcc ttc gat gag gac agc agc gat gcc tta tcc ccc 925 Ala Asp Ser Ser SerPhe Asp Glu Asp Ser Ser Asp Ala Leu Ser Pro 110 115 120 125 gag cag cctgcc agc cat gag tcc cag ggt tct gtg ccg tca ccc ctg 973 Glu Gln Pro AlaSer His Glu Ser Gln Gly Ser Val Pro Ser Pro Leu 130 135 140 gag gcc cgagtc agc gaa cca ctg ctc agt gcc acc tct gca tcc ccc 1021 Glu Ala Arg ValSer Glu Pro Leu Leu Ser Ala Thr Ser Ala Ser Pro 145 150 155 acc cag gttgtg tct caa ctt ccg atg ggc cgg gat tcc aga gaa atg 1069 Thr Gln Val ValSer Gln Leu Pro Met Gly Arg Asp Ser Arg Glu Met 160 165 170 ctt ttc ctggca gag cag cct cct ctg cct ccc cca cct ctg ctg cct 1117 Leu Phe Leu AlaGlu Gln Pro Pro Leu Pro Pro Pro Pro Leu Leu Pro 175 180 185 ccc agc ctcacc aat gga acc act atc ccc act gcc aag tcc acc ccc 1165 Pro Ser Leu ThrAsn Gly Thr Thr Ile Pro Thr Ala Lys Ser Thr Pro 190 195 200 205 aca ctcatt aag caa agc caa ccc aag tct gcc agt gag aag tca cag 1213 Thr Leu IleLys Gln Ser Gln Pro Lys Ser Ala Ser Glu Lys Ser Gln 210 215 220 cgc agcaag aag gcc aag gag ctg aag cca aag gtg aag aag ctc aag 1261 Arg Ser LysLys Ala Lys Glu Leu Lys Pro Lys Val Lys Lys Leu Lys 225 230 235 tac caccag tac atc ccc ccg gac cag aag cag gac agg ggg gca ccc 1309 Tyr His GlnTyr Ile Pro Pro Asp Gln Lys Gln Asp Arg Gly Ala Pro 240 245 250 ccc atggac tca tcc tac gcc aag atc ctg cag cag cag cag ctc ttc 1357 Pro Met AspSer Ser Tyr Ala Lys Ile Leu Gln Gln Gln Gln Leu Phe 255 260 265 ctc cagctg cag atc ctc aac cag cag cag cag cag cac cac aac tac 1405 Leu Gln LeuGln Ile Leu Asn Gln Gln Gln Gln Gln His His Asn Tyr 270 275 280 285 caggcc atc ctg cct gcc ccg cca aag tca gca ggc gag gcc ctg gga 1453 Gln AlaIle Leu Pro Ala Pro Pro Lys Ser Ala Gly Glu Ala Leu Gly 290 295 300 agcagc ggg acc ccc cca gta cgc agc ctc tcc act acc aat agc agc 1501 Ser SerGly Thr Pro Pro Val Arg Ser Leu Ser Thr Thr Asn Ser Ser 305 310 315 tccagc tcg ggc gcc cct ggg ccc tgt ggg ctg gca cgt cag aac agc 1549 Ser SerSer Gly Ala Pro Gly Pro Cys Gly Leu Ala Arg Gln Asn Ser 320 325 330 acctca ctg act ggc aag ccg gga gcc ctg ccg gcc aac ctg gac gac 1597 Thr SerLeu Thr Gly Lys Pro Gly Ala Leu Pro Ala Asn Leu Asp Asp 335 340 345 atgaag gtg gca gag ctg aag cag gag ctg aag ttg cga tca ctg cct 1645 Met LysVal Ala Glu Leu Lys Gln Glu Leu Lys Leu Arg Ser Leu Pro 350 355 360 365gtc tcg ggc acc aaa act gag ctg att gag cgc ctt cga gcc tat caa 1693 ValSer Gly Thr Lys Thr Glu Leu Ile Glu Arg Leu Arg Ala Tyr Gln 370 375 380gac caa atc agc cct gtg cca gga gcc ccc aag gcc cct gcc gcc acc 1741 AspGln Ile Ser Pro Val Pro Gly Ala Pro Lys Ala Pro Ala Ala Thr 385 390 395tct atc ctg cac aag gct ggc gag gtg gtg gta gcc ttc cca gcg gcc 1789 SerIle Leu His Lys Ala Gly Glu Val Val Val Ala Phe Pro Ala Ala 400 405 410cgg ctg agc acg ggg cca gcc ctg gtg gca gca ggc ctg gct cca gct 1837 ArgLeu Ser Thr Gly Pro Ala Leu Val Ala Ala Gly Leu Ala Pro Ala 415 420 425gag gtg gtg gtg gcc acg gtg gcc agc agt ggg gtg gtg aag ttt ggc 1885 GluVal Val Val Ala Thr Val Ala Ser Ser Gly Val Val Lys Phe Gly 430 435 440445 agc acg ggc tcc acg ccc ccc gtg tct ccc acc ccc tcg gag cgc tca 1933Ser Thr Gly Ser Thr Pro Pro Val Ser Pro Thr Pro Ser Glu Arg Ser 450 455460 ctg ctc agc acg ggc gat gaa aac tcc acc ccc ggg gac acc ttt ggt 1981Leu Leu Ser Thr Gly Asp Glu Asn Ser Thr Pro Gly Asp Thr Phe Gly 465 470475 gag atg gtg aca tca cct ctg acg cag ctg acc ctg cag gcc tcg cca 2029Glu Met Val Thr Ser Pro Leu Thr Gln Leu Thr Leu Gln Ala Ser Pro 480 485490 ctg cag atc ctc gtg aag gag gag ggc ccc cgg gcc ggg tcc tgt tgc 2077Leu Gln Ile Leu Val Lys Glu Glu Gly Pro Arg Ala Gly Ser Cys Cys 495 500505 ctg agc cct ggg ggg cgg gcg gag cta gag ggg cgc gac aag gac cag 2125Leu Ser Pro Gly Gly Arg Ala Glu Leu Glu Gly Arg Asp Lys Asp Gln 510 515520 525 atg ctg cag gag aaa gac aag cag atc gag gcg ctg acg cgc atg ctc2173 Met Leu Gln Glu Lys Asp Lys Gln Ile Glu Ala Leu Thr Arg Met Leu 530535 540 cgg cag aag cag cag ctg gtg gag cgg ctc aag ctg cag ctg gag cag2221 Arg Gln Lys Gln Gln Leu Val Glu Arg Leu Lys Leu Gln Leu Glu Gln 545550 555 gag aag cga gcc cag cag ccc gcc ccc gcc ccc gcc ccc ctc ggc acc2269 Glu Lys Arg Ala Gln Gln Pro Ala Pro Ala Pro Ala Pro Leu Gly Thr 560565 570 ccc gtg aag cag gag aac agc ttc tcc agc tgc cag ctg agc cag cag2317 Pro Val Lys Gln Glu Asn Ser Phe Ser Ser Cys Gln Leu Ser Gln Gln 575580 585 ccc ctg ggc ccc gct cac cca ttc aac ccc agc ctg gcg gcc cca gcc2365 Pro Leu Gly Pro Ala His Pro Phe Asn Pro Ser Leu Ala Ala Pro Ala 590595 600 605 acc aac cac ata gac cct tgt gct gtg gcc ccg ggg ccc ccg tccgtg 2413 Thr Asn His Ile Asp Pro Cys Ala Val Ala Pro Gly Pro Pro Ser Val610 615 620 gtg gtg aag cag gaa gcc ttg cag cct gag ccc gag ccg gtc cccgcc 2461 Val Val Lys Gln Glu Ala Leu Gln Pro Glu Pro Glu Pro Val Pro Ala625 630 635 ccc cag ttg ctt ctg ggg cct cag ggc ccc agc ctc atc aag ggggtt 2509 Pro Gln Leu Leu Leu Gly Pro Gln Gly Pro Ser Leu Ile Lys Gly Val640 645 650 gca cct ccc acc ctc atc acc gac tcc aca ggg acc cac ctt gtcctc 2557 Ala Pro Pro Thr Leu Ile Thr Asp Ser Thr Gly Thr His Leu Val Leu655 660 665 acc gtg acc aat aag aat gca gac agc cct ggc ctg tcc agt gggagc 2605 Thr Val Thr Asn Lys Asn Ala Asp Ser Pro Gly Leu Ser Ser Gly Ser670 675 680 685 ccc cag cag ccc tcg tcc cag cct ggc tct cca gcg cct gccccc tct 2653 Pro Gln Gln Pro Ser Ser Gln Pro Gly Ser Pro Ala Pro Ala ProSer 690 695 700 gcc cag atg gac ctg gag cac cca ctg cag ccc ctc ttt gggacc ccc 2701 Ala Gln Met Asp Leu Glu His Pro Leu Gln Pro Leu Phe Gly ThrPro 705 710 715 act tct ctg ctg aag aag gaa cca cct ggc tat gag gaa gccatg agc 2749 Thr Ser Leu Leu Lys Lys Glu Pro Pro Gly Tyr Glu Glu Ala MetSer 720 725 730 cag cag ccc aaa cag cag gaa aat ggt tcc tca agc cag cagatg gac 2797 Gln Gln Pro Lys Gln Gln Glu Asn Gly Ser Ser Ser Gln Gln MetAsp 735 740 745 gac ctg ttt gac att ctc att cag agc gga gaa att tca gcagat ttc 2845 Asp Leu Phe Asp Ile Leu Ile Gln Ser Gly Glu Ile Ser Ala AspPhe 750 755 760 765 aag gag ccg cca tcc ctg cca ggg aag gag aag cca tccccg aag aca 2893 Lys Glu Pro Pro Ser Leu Pro Gly Lys Glu Lys Pro Ser ProLys Thr 770 775 780 gtc tgt ggg tcc ccc ctg gca gca cag cca tca cct tctgct gag ctc 2941 Val Cys Gly Ser Pro Leu Ala Ala Gln Pro Ser Pro Ser AlaGlu Leu 785 790 795 ccc cag gct gcc cca cct cct cca ggc tca ccc tcc ctccct gga cgc 2989 Pro Gln Ala Ala Pro Pro Pro Pro Gly Ser Pro Ser Leu ProGly Arg 800 805 810 ctg gag gac ttc ctg gag agc agc acg ggg ctg ccc ctgctg acc agt 3037 Leu Glu Asp Phe Leu Glu Ser Ser Thr Gly Leu Pro Leu LeuThr Ser 815 820 825 ggg cat gac ggg cca gag ccc ctt tcc ctc att gac gacctc cat agc 3085 Gly His Asp Gly Pro Glu Pro Leu Ser Leu Ile Asp Asp LeuHis Ser 830 835 840 845 cag atg ctg agc agc act gcc atc ctg gac cac cccccg tca ccc atg 3133 Gln Met Leu Ser Ser Thr Ala Ile Leu Asp His Pro ProSer Pro Met 850 855 860 gac acc tcg gaa ttg cac ttt gtt cct gag ccc agcagc acc atg ggc 3181 Asp Thr Ser Glu Leu His Phe Val Pro Glu Pro Ser SerThr Met Gly 865 870 875 ctg gac ctg gct gat ggc cac ctg gac agc atg gactgg ctg gag ctg 3229 Leu Asp Leu Ala Asp Gly His Leu Asp Ser Met Asp TrpLeu Glu Leu 880 885 890 tcg tca ggt ggt ccc gtg ctg agc cta gcc ccc ctcagc acc aca gcc 3277 Ser Ser Gly Gly Pro Val Leu Ser Leu Ala Pro Leu SerThr Thr Ala 895 900 905 ccc agc ctc ttc tcc aca gac ttc ctc gat ggc catgat ttg cag ctg 3325 Pro Ser Leu Phe Ser Thr Asp Phe Leu Asp Gly His AspLeu Gln Leu 910 915 920 925 cac tgg gat tcc tgc ttg tag ctctctggctcaagacgggg tggggaaggg 3376 His Trp Asp Ser Cys Leu 930 gctgggagccagggtactcc aatgcgtggc tctcctgcgt gattcggcct ctccacatgg 3436 ttgtgagtcttgacaatcac agcccctgct ttttcccttc cctgggaggc tagaacagag 3496 aagcccttactcctggttca gtgccacgca gggcagagga gagcagctgt caagaagcag 3556 ccctggctctcacgctgggg ttttggacac acggtcaggg tcagggccat ttcagcttga 3616 cctccttttttgaggtcagg gggcactgtc tgtctggcta caatttggct aaggtaggtg 3676 aagcctggccaggcgggagg cttctcttct gacccagggc tgagacaggt taaggggtga 3736 atctccttcctttctctccc tgctttgctg tgaagggaga aattagcctg ggcctctacc 3796 ccctattccctgtgtctgcc aaccccagga tcccagggct ccctgccatt ttagtgtctt 3856 ggtgtagtgtaaccatttag tggttggtgg caacaatttt atgtacaggt gtatatacct 3916 ctatattatatatcgacata catatatatt tttggggggg ggcggacagg agatgggtgc 3976 aactccctcccatcctactc tcacagaagg gcctggatgc aaggttaccc ttgagctgtg 4036 tgccacagtctggtgcccag tctggcatgc agctacccag gcccacccat cacgtgtgat 4096 tgacatgtaggtaccctgcc acggcctatg ccccacctgc cctgcttcct ggctccttat 4156 cagtgccatgagggcagagg tgctacctgg ccttcctgcc aggagctctc cacccactca 4216 cattccgtccccgccgcctc actgcagcca gcgtggtcct aggacaggag gagcttcggg 4276 cccagcttcaccctgcggtg gggctgaggg gtggccatct cctgccctgg ggccactggc 4336 ttcacattctgggctgactc ataggggagt aggggtggag tcaccaaaac cagtgctggg 4396 acaaagatggggaaggtgtg tgaacttttt aaaataaaca caaaaacaca g 4447 14 931 PRT Homosapiens 14 Met Pro Pro Leu Lys Ser Pro Ala Ala Phe His Glu Gln Arg ArgSer 1 5 10 15 Leu Glu Arg Ala Arg Thr Glu Asp Tyr Leu Lys Arg Lys IleArg Ser 20 25 30 Arg Pro Glu Arg Ser Glu Leu Val Arg Met His Ile Leu GluGlu Thr 35 40 45 Ser Ala Glu Pro Ser Leu Gln Ala Lys Gln Leu Lys Leu LysArg Ala 50 55 60 Arg Leu Ala Asp Asp Leu Asn Glu Lys Ile Ala Gln Arg ProGly Pro 65 70 75 80 Met Glu Leu Val Glu Lys Asn Ile Leu Pro Val Glu SerSer Leu Lys 85 90 95 Glu Ala Ile Ile Val Gly Gln Val Asn Tyr Pro Lys ValAla Asp Ser 100 105 110 Ser Ser Phe Asp Glu Asp Ser Ser Asp Ala Leu SerPro Glu Gln Pro 115 120 125 Ala Ser His Glu Ser Gln Gly Ser Val Pro SerPro Leu Glu Ala Arg 130 135 140 Val Ser Glu Pro Leu Leu Ser Ala Thr SerAla Ser Pro Thr Gln Val 145 150 155 160 Val Ser Gln Leu Pro Met Gly ArgAsp Ser Arg Glu Met Leu Phe Leu 165 170 175 Ala Glu Gln Pro Pro Leu ProPro Pro Pro Leu Leu Pro Pro Ser Leu 180 185 190 Thr Asn Gly Thr Thr IlePro Thr Ala Lys Ser Thr Pro Thr Leu Ile 195 200 205 Lys Gln Ser Gln ProLys Ser Ala Ser Glu Lys Ser Gln Arg Ser Lys 210 215 220 Lys Ala Lys GluLeu Lys Pro Lys Val Lys Lys Leu Lys Tyr His Gln 225 230 235 240 Tyr IlePro Pro Asp Gln Lys Gln Asp Arg Gly Ala Pro Pro Met Asp 245 250 255 SerSer Tyr Ala Lys Ile Leu Gln Gln Gln Gln Leu Phe Leu Gln Leu 260 265 270Gln Ile Leu Asn Gln Gln Gln Gln Gln His His Asn Tyr Gln Ala Ile 275 280285 Leu Pro Ala Pro Pro Lys Ser Ala Gly Glu Ala Leu Gly Ser Ser Gly 290295 300 Thr Pro Pro Val Arg Ser Leu Ser Thr Thr Asn Ser Ser Ser Ser Ser305 310 315 320 Gly Ala Pro Gly Pro Cys Gly Leu Ala Arg Gln Asn Ser ThrSer Leu 325 330 335 Thr Gly Lys Pro Gly Ala Leu Pro Ala Asn Leu Asp AspMet Lys Val 340 345 350 Ala Glu Leu Lys Gln Glu Leu Lys Leu Arg Ser LeuPro Val Ser Gly 355 360 365 Thr Lys Thr Glu Leu Ile Glu Arg Leu Arg AlaTyr Gln Asp Gln Ile 370 375 380 Ser Pro Val Pro Gly Ala Pro Lys Ala ProAla Ala Thr Ser Ile Leu 385 390 395 400 His Lys Ala Gly Glu Val Val ValAla Phe Pro Ala Ala Arg Leu Ser 405 410 415 Thr Gly Pro Ala Leu Val AlaAla Gly Leu Ala Pro Ala Glu Val Val 420 425 430 Val Ala Thr Val Ala SerSer Gly Val Val Lys Phe Gly Ser Thr Gly 435 440 445 Ser Thr Pro Pro ValSer Pro Thr Pro Ser Glu Arg Ser Leu Leu Ser 450 455 460 Thr Gly Asp GluAsn Ser Thr Pro Gly Asp Thr Phe Gly Glu Met Val 465 470 475 480 Thr SerPro Leu Thr Gln Leu Thr Leu Gln Ala Ser Pro Leu Gln Ile 485 490 495 LeuVal Lys Glu Glu Gly Pro Arg Ala Gly Ser Cys Cys Leu Ser Pro 500 505 510Gly Gly Arg Ala Glu Leu Glu Gly Arg Asp Lys Asp Gln Met Leu Gln 515 520525 Glu Lys Asp Lys Gln Ile Glu Ala Leu Thr Arg Met Leu Arg Gln Lys 530535 540 Gln Gln Leu Val Glu Arg Leu Lys Leu Gln Leu Glu Gln Glu Lys Arg545 550 555 560 Ala Gln Gln Pro Ala Pro Ala Pro Ala Pro Leu Gly Thr ProVal Lys 565 570 575 Gln Glu Asn Ser Phe Ser Ser Cys Gln Leu Ser Gln GlnPro Leu Gly 580 585 590 Pro Ala His Pro Phe Asn Pro Ser Leu Ala Ala ProAla Thr Asn His 595 600 605 Ile Asp Pro Cys Ala Val Ala Pro Gly Pro ProSer Val Val Val Lys 610 615 620 Gln Glu Ala Leu Gln Pro Glu Pro Glu ProVal Pro Ala Pro Gln Leu 625 630 635 640 Leu Leu Gly Pro Gln Gly Pro SerLeu Ile Lys Gly Val Ala Pro Pro 645 650 655 Thr Leu Ile Thr Asp Ser ThrGly Thr His Leu Val Leu Thr Val Thr 660 665 670 Asn Lys Asn Ala Asp SerPro Gly Leu Ser Ser Gly Ser Pro Gln Gln 675 680 685 Pro Ser Ser Gln ProGly Ser Pro Ala Pro Ala Pro Ser Ala Gln Met 690 695 700 Asp Leu Glu HisPro Leu Gln Pro Leu Phe Gly Thr Pro Thr Ser Leu 705 710 715 720 Leu LysLys Glu Pro Pro Gly Tyr Glu Glu Ala Met Ser Gln Gln Pro 725 730 735 LysGln Gln Glu Asn Gly Ser Ser Ser Gln Gln Met Asp Asp Leu Phe 740 745 750Asp Ile Leu Ile Gln Ser Gly Glu Ile Ser Ala Asp Phe Lys Glu Pro 755 760765 Pro Ser Leu Pro Gly Lys Glu Lys Pro Ser Pro Lys Thr Val Cys Gly 770775 780 Ser Pro Leu Ala Ala Gln Pro Ser Pro Ser Ala Glu Leu Pro Gln Ala785 790 795 800 Ala Pro Pro Pro Pro Gly Ser Pro Ser Leu Pro Gly Arg LeuGlu Asp 805 810 815 Phe Leu Glu Ser Ser Thr Gly Leu Pro Leu Leu Thr SerGly His Asp 820 825 830 Gly Pro Glu Pro Leu Ser Leu Ile Asp Asp Leu HisSer Gln Met Leu 835 840 845 Ser Ser Thr Ala Ile Leu Asp His Pro Pro SerPro Met Asp Thr Ser 850 855 860 Glu Leu His Phe Val Pro Glu Pro Ser SerThr Met Gly Leu Asp Leu 865 870 875 880 Ala Asp Gly His Leu Asp Ser MetAsp Trp Leu Glu Leu Ser Ser Gly 885 890 895 Gly Pro Val Leu Ser Leu AlaPro Leu Ser Thr Thr Ala Pro Ser Leu 900 905 910 Phe Ser Thr Asp Phe LeuAsp Gly His Asp Leu Gln Leu His Trp Asp 915 920 925 Ser Cys Leu 930 1524 DNA synthetic construct 15 aggctggact caacaggaag gatg 24 16 23 DNAsynthetic construct 16 cctggcccgc tccaagctcc ttc 23 17 23 DNA syntheticconstruct 17 ctgctcatga aatgcggctg gac 23 18 22 DNA synthetic construct18 tctcccagca gttcctggat tc 22 19 24 DNA synthetic construct 19ctggctctct tcagcttcag ctgc 24 20 22 DNA synthetic construct 20tctccttgca gcccgagctg ac 22 21 24 DNA synthetic construct 21 ccatcagaggcccatgtaaa ctcc 24 22 24 DNA synthetic construct 22 gttacacacggactttagga cgca 24 23 23 DNA synthetic construct 23 catgcttctactgaagaaca gca 23 24 21 DNA synthetic construct 24 aatccaggaa ctgctgggaga 21 25 20 DNA synthetic construct 25 actggttgaa cagcggatga 20 26 20 DNAsynthetic construct 26 aactgtgagc gcacgtttga 20 27 21 DNA syntheticconstruct 27 gtcctaaagt ccgtgtgtaa c 21 28 21 DNA synthetic construct 28ggtcagtgga ctcctttctc c 21 29 22 DNA synthetic construct 29 gctgttgccaatgaactgca gg 22

1. An isolated fusion protein associated with acute megakaryoblasticleukemia (AMKL) comprising an amino acid sequence selected from thegroup consisting of: (a) the amino acid sequence set forth in SEQ ID NO:2; (b) an amino acid sequence having at least 90% amino acid sequenceidentity to the amino acid sequence of (a); (c) the amino acid sequenceencoded by the nucleotide sequence set forth in SEQ ID NO: 1; (d) theamino acid sequence set forth in SEQ ID NO: 4; (e) an amino acidsequence having at least 90% amino acid sequence identity to the aminoacid sequence of (d): (f) the amino acid sequence encoded by thenucleotide sequence set forth in SEQ ID NO: 3; (g) the amino acidsequence set forth in SEQ ID NO: 6; (h) an amino acid sequence having atleast 90% amino acid sequence identity to the amino acid sequence of(g); and (i) the amino acid sequence encoded by the nucleotide sequenceset forth in SEQ ID NO:
 5. 2. An isolated nucleotide molecule comprisinga nucleotide sequence selected from the group consisting of: (a) anucleotide sequence encoding the fusion protein having the amino acidsequence set forth in SEQ ID NO: 2; (b) the nucleotide sequence setforth in SEQ ID NO: 1; (c) a nucleotide sequence having at least 90%nucleotide sequence identity to the nucleotide sequence of (b); (d) anucleotide sequence encoding the fusion protein having the amino acidsequence set forth in SEQ ID NO: 4; (e) the nucleotide sequence setforth in SEQ ID NO: 3; and (f) a nucleotide sequence having at least 90%nucleotide sequence identity to the nucleotide sequence of (e); (g) anucleotide sequence encoding the fusion protein having the amino acidsequence set forth in SEQ ID NO: 6; (h) the nucleotide sequence setforth in SEQ ID NO: 5; and (i) a nucleotide sequence having at least 90%nucleotide sequence identity to the nucleotide sequence of (h). 3-6.(canceled)
 7. An antibody that specifically binds to an fusion proteincomprising an amino acid sequence selected from the group consisting of:(a) the amino acid sequence set forth in SEQ ID NO: 2; (b) an amino acidsequence having at least 90% amino acid sequence identity to the aminoacid sequence of (a); (c) the amino acid sequence encoded by thenucleotide sequence set forth in SEQ ID NO: 1; (d) the amino acidsequence set forth in SEQ ID NO: 4; (e) an amino acid sequence having atleast 90% amino acid sequence identity to the amino acid sequence of(d); (f) the amino acid sequence encoded by the nucleotide sequence setforth in SEQ ID NO: 3; (g) the amino acid sequence set forth in SEQ IDNO: 6; (h) an amino acid sequence having at least 90% amino acidsequence identity to the amino acid sequence of (g); and (i) the aminoacid sequence encoded by the nucleotide sequence set forth in SEQ ID NO:5. 8-9. (canceled)
 10. A non-human transgenic animal that has beenaltered to express a gene encoding an fusion protein comprising an aminoacid sequence selected from the group consisting of: (a) the amino acidsequence set forth in SEQ ID NO: 2; (b) an amino acid sequence having atleast 90% amino acid sequence identity to the amino acid sequence of(a); (c) the amino acid sequence encoded by the nucleotide sequence setforth in SEQ ID NO: 1; (d) the amino acid sequence set forth in SEQ IDNO: 4; (e) an amino acid sequence having at least 90% amino acidsequence identity to the amino acid sequence of (d); (f) the amino acidsequence encoded by the nucleotide sequence set forth in SEQ ID NO: 3;(g) the amino acid sequence set forth in SEQ ID NO: 6; (h) an amino acidsequence having at least 90% amino acid sequence identity to the aminoacid sequence of (g); and (i) the amino acid sequence encoded by thenucleotide sequence set forth in SEQ ID NO:
 5. 11-12. (canceled)
 13. Amethod of identifying an agent capable of binding to a fusion proteinhaving an amino acid sequence selected from the group consisting of SEQID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, and variants thereof, comprisingthe steps of: (a) contacting a candidate agent with said fusion protein;and (b) determining whether said candidate agent binds said fusionprotein.
 14. The method of claim 13 further comprising the additionalsteps of: (c) determining whether said candidate agent binds to an RBM15protein having an amino acid sequence selected from the group consistingof SEQ ID NOS: 8, 10 and 12, and variants thereof; and (d) determiningwhether said candidate agent binds to an MKL1 protein having the aminoacid sequence set forth in SEQ ID NO: 14, or variant thereof; wherein acandidate agent which binds to said fusion protein but does not bindsignificantly to said RBM15 protein or said MKL1protein is identified asan agent which selectively binds to said RBM15-MKL1 fusion protein.15-18. (canceled)
 19. A method for detecting the t(1;22) chromosomalrearrangement associated with AMKL comprising: (a) obtaining a samplecomprising RNA; (b) performing reverse transcriptase-polymerase chainreaction (RT-PCR) amplification with said sample so as to amplify a cDNAcomprising at least a portion of nucleotide sequence selected from thegroup consisting of SEQ ID NO: 1, 3, and 5 spanning the fusion regionbetween the RBM15 and MKL1 coding sequences; and (c) detecting thepresence of said cDNA, wherein the presence of said DNA indicates thepresence of said rearrangement in said sample or in cells from whichsaid sample is derived.
 20. A method for detecting the t(1;22)chromosomal rearrangement associated with AMKL comprising: (a)hybridizing a probe labeled with a detection reagent to a samplecomprising a human chromosome band 1p13 and a human chromosome band22q13, wherein: said probe comprises nucleotide sequences from a wildtype chromosome 1 or 22; said probe comprises nucleotide sequences thatare known to occur in the proximity of the t(1,22) breakpoint; and saidprobe comprises at least one polynucleotide; (b) detecting said probe bydetecting said detection reagent; and (c) determining the location ofsaid probe, wherein a signal that localizes to the same chromosomeindicates the presence of t(1;22) chromosomal rearrangement.
 21. Themethod of claim of 20, wherein said probe comprises nucleotide sequencesthat are known to occur within 10 kb of the t(1,22) breakpoint.
 22. Amethod for detecting the t(1;22) chromosomal rearrangement associatedwith acute megakaryoblastic leukemia comprising: (a) hybridizing a firstprobe labeled with a first detection reagent to a sample comprising ahuman chromosome band 1p13 and a human chromosome band 22q13; whereinsaid first probe comprises at least one polynucleotide, and said probeis capable of hybridization to human chromosome band 1p13 in theproximity of the t(1;22) chromosomal rearrangement; (b) hybridizing adifferently labeled second probe labeled with a second detection reagentto said sample, wherein: said second probe comprises at least onepolynucleotide, said probe is capable of hybridization to humanchromosome band 22q13 in the proximity of the t(1;22) chromosomalrearrangement; and (c) detecting said first probe and said second probeby detecting said first detection reagent and said second detectionreagent; and (d) determining the location of said first probe relativeto said second probe, wherein the appearance of said first probe andsaid second probe as paired signals that localize to one humanchromosome indicates the presence of a t(1;22) chromosomal rearrangementin said sample.
 23. The method of claim of 22, wherein said first andsecond probes comprise nucleotide sequences that are known to occurwithin 10 kb of the t(1,22) breakpoint.
 24. A method of screening foragents capable of inhibiting the activity of a fusion protein arisingfrom the t(1;22) chromosomal rearrangement associated with AMKLcomprising: (a) expressing in a eukaryotic cell at least one fusionprotein, where said fusion protein is selected from the group consistingof RBM15-MK1, MK1-RBM15_(S), MK1-RBM15_(S+AE), and variants thereof; (b)administering said agent to said cell; and (c) monitoring said cell fora suppression or elimination of an adverse phenotype associated withexpression of the fusion protein selected in step (a); wherein an agentwhich suppresses or eliminates said adverse phenotype is identified asan inhibitor of the fusion protein selected in step (a).
 25. A method ofscreening for agents capable of selectively inhibiting the activity of afusion protein arising from the t(1;22) chromosomal rearrangementassociated with AMKL comprising: (a) expressing in a first eukaryoticcell at least one fusion protein, where said fusion protein is selectedfrom the group consisting of RBM15-MKL1, MKL1-RBM15_(S),MKL1-RBM15_(S+AE), and variants thereof; (b) administering said agent tosaid first eukaryotic cell; (c) monitoring said first eukaryotic cellfor a suppression or elimination of an adverse phenotype associated withexpression of the fusion protein selected in step (a); (d) expressing ina second eukaryotic cell an RBM15 protein; (e) monitoring said secondeukaryotic cell for a suppression or elimination of a phenotypeassociated with expression of RBM15 protein; (f) expressing in a thirdeukaryotic cell an MKL1protein; and (g) monitoring said third eukaryoticcell for a suppression or elimination of a phenotype associated withexpression of MKL1protein; wherein an agent which suppresses oreliminates the adverse phenotype of the fusion protein selected in step(a) but does not suppress or eliminate the phenotype of the RBM15protein or the MKL1protein is identified as a selective inhibitor of thefusion protein selected in step (a).
 26. The method of claim 13, whereinsaid fusion protein is a RBM15-MLK1 fusion protein having the amino acidsequence set forth in SEQ ID NO: 2, or variant thereof.
 27. The methodof claim 14, wherein said fusion protein is a RBM15-MLK1 fusion proteinhaving the amino acid sequence set forth in SEQ ID NO: 2, or variantthereof.
 28. The method of claim 13, wherein the fusion protein is anMKL1-RBM15_(S) fusion protein having the amino acid sequence set forthin SEQ ID NO: 4, or variant thereof.
 29. The method of claim 14, whereinthe fusion protein is an MKL1-RBM15_(S) fusion protein having the aminoacid sequence set forth in SEQ ID NO: 4, or variant thereof.
 30. Themethod of claim 13, wherein the fusion protein is an MKL1-RBM15_(S+AE)fusion protein having the amino acid sequence set forth in SEQ ID NO: 6,or variant thereof.
 31. The method of claim 14, wherein the fusionprotein is an MKL1-RBM15_(S+AE) fusion protein having the amino acidsequence set forth in SEQ ID NO: 6, or variant thereof.